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STUDIES AVAILABLE OF VIDARIKAND

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1. The effect of Pueraria mirifica on cytologic and urodynamic findings in ovariectomized rats.

Manonai J, Seif C, Böhler G, Jünemann KP.

From the Departments of 1Obstetrics and Gynaecology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand; and 2Urology and Pediatric Urology, University Hospital Schleswig-Holstein, Campus Kiel, Germany.

OBJECTIVE:: To evaluate the effect of Pueraria mirifica on vaginal and urethral cytology, bladder pressure and capacity, residual urine, and leak point pressure in ovariectomized rats. DESIGN:: Seventy-two adult, ovariectomized, female Sprague-Dawley rats were placed into one of four groups: control, estradiol, or 100 or 1,000 mg/kg of Pueraria mirifica (PM-100 and PM-1000, respectively). The vaginal and urethral smears were checked after 30 days of ovariectomy at pretreatment and at day 28 of treatment. A single cystometry, defined as the micturition interval, filling pressure, threshold pressure, micturition pressure, and voided volume, was performed. Peak bladder pressure was calculated for each leak point pressure measured at half bladder capacity by slowly and manually increasing abdominal pressure until a leak occurred, at which point external pressure was rapidly released. Leak point pressure was tested three times per rat. RESULTS:: After 28 days of treatment, the estradiol, PM-100, and PM-1000 groups had significantly higher numbers of vaginal and urethral superficial cells compared with the control group (P < 0.05). Regarding the urodynamic parameters, the threshold pressure, micturition pressure, and leak point pressure were higher in the estradiol, PM-100, and PM-1000 groups compared with the control group (P < 0.05). The control, PM-100, and PM-1000 groups had the same values for micturition interval, bladder capacity, voided volume, and residual volume (P > 0.05) but lower values compared with the estradiol group (P < 0.05). CONCLUSIONS:: Pueraria mirifica 100 and 1,000 mg/kg/day showed an estrogen-like effect on the vaginal and urethral epithelium of ovariectomized rats. They did not change bladder capacity and residual urine volume but increased leak point pressure according to urodynamic study.

PMID: 19098688 [PubMed - as supplied by publisher]

2. An investigation on the germination of seeds of Kwao Kreu Kao (Pueraria candollei Grah. ex Benth) as affected by both water soakings and hot air oven treatments.

Benjawan C, Chutichudet P, Kaewsit S.

Department of Agricultural Technology, Faculty of Technology, Mahasarakham University, Mahasarakham 44000, Thailand.

This experiment was carried out at the Faculty of Technology, Mahasarakham University, Mahasarakham 4400, Thailand to investigate effects of different water-soaking durations and hot air oven treatments on the germination of seeds of Kwao Kreu Kao (Pueraria candollei Grah. ex Benth) plants. The experiment was laid in a Randomised Complete Block Design with four replications. The results showed that after tested for Electrical Conductivity (EC) values for cracking of seeds, all seeds being used were at a normal condition (with an average EC value of 28.56 microS cm(-1) g(-1)) and all seeds were ready for germination. Strength on impermeability of seeds declined after soaking in water for 10 h and onwards then the treated seeds had increased in weights. However, after treated under hot air oven, dry weights of all seeds became similar. Germination percentage of all treatments was most rapid during the first three weeks of germination period and later slowly increased with time. At day 91 after sowing, T2 gave the highest percentage of germination (52%) and the lowest was found with T1 (control) with 31.25%. Again at day 91 after sowing, T2 gave the highest mean value of plant numbers (16.38) and the lowest was found with T1 (7.28). Numbers of abnormal seedlings determined at day 63 after sowing were lowest with T2 (6.25%) and worst with T4 (20.14%). Again at day 63 after sowing, plant height was significantly tallest with T2 (3.88 cm) and the lowest was found with T4 (2.71 cm). Numbers of leaves were not significantly different among the treated plants reaching a highest value of 11.25 leaves plant(-1) for T3. It may be concluded that T2 was the best treatment for use in germinating seeds of Pueraria candollei Grah. ex Benth plants. Further improvements on longevity and high percentage of germination of seeds were discussed and suggested.

Publication Types: PMID: 19086573 [PubMed - in process]

3. Pueraria mirifica phytoestrogens improve dyslipidemia in postmenopausal women probably by activating estrogen receptor subtypes.

Okamura S, Sawada Y, Satoh T, Sakamoto H, Saito Y, Sumino H, Takizawa T, Kogure T, Chaichantipyuth C, Higuchi Y, Ishikawa T, Sakamaki T
.

Department of Medical Informatics and Decision Sciences, School of Medicine, Gunma University, Maebashi, Japan.

Impaired lipid metabolism is an important health problem in postmenopausal women with insufficient estrogens, because dyslipidemia is a risk factor for development of atherosclerosis and the incidence of cardiovascular disease markedly increases after menopause. Pueraria mirifica (PM), a Thai herb, has been noticed as a source of phytoestrogens, estrogen-mimicking plant compounds. However, the clinical effects of PM on lipid metabolism and the underlying molecular mechanisms remain undetermined. Therefore, we examined the effects of PM on serum lipid parameters in a randomized, double-blind, placebo-controlled clinical trial. Nineteen postmenopausal women were randomly assigned to receive oral administration of PM powder or placebo. After 2 months of treatment, the PM group showed a significant increase in serum concentrations of high-density lipoprotein (HDL) cholesterol and apolipoprotein (apo) A-1 (34% and 40%, respectively), and a significant decrease in low-density lipoprotein (LDL) cholesterol and apo B (17% and 9%, respectively), compared with baseline measurements. Moreover, significant decreases were observed in the ratios of LDL cholesterol to HDL cholesterol (37%) and apo B to apo A-1 (35%). Next, we determined the effects of PM phytoestrogens on the activation of estrogen receptor (ER)-mediated transactivation by transient expression assays of a reporter gene in cultured cells. Among PM phytoestrogens, miroestrol and coumestrol enhanced both ERalpha- and ERbeta-mediated transactivation, whereas other phytoestrogens, including daidzein and genistein, preferentially enhanced ERbeta-mediated transactivation. In conclusion, PM has a beneficial effect on lipid metabolism in postmenopausal women, which may result from the activation of gene transcription through selective binding of phytoestrogens to ERalpha and ERbeta.

Publication Types: PMID: 19060449 [PubMed - in process]

4. Plant extracts as natural amoebicidal agents.

Derda M, Hadaś E, Thiem B.

Department of Biology and Medical Parasitology, Poznan University of Medical Sciences, 10 Fredry Street, Poznań, 61-701, Poland, mderda@ump.edu.pl.

Strains of Acanthamoeba sp. constitute a factor contributing to the occurrence of chronic granulomatous amoebic encephalitis, keratitis, pneumonia, as well as inflammations of other organs. Treatment of these diseases is very difficult and not always effective. A majority of these infections have been fatal. The aim of our study was to examine the amoebicidal or amoebistatic activity of plant extracts from Rubus chamaemorus, Pueraria lobata, Solidago virgaurea and Solidago graminifolia. For the purpose of isolation of pharmacologically active substances, we used the aboveground parts of plants, together with flowers, roots and leaves. It was established that extracts from S. virgauera, P. lobata and R. chamaemorus displayed chemotherapeutic properties in vitro in concentrations of approximately 0.01-0.05 mg extract/mL, i.e. in concentrations of 0.350 mug/mL expressed in ellagic acid for R. chamaemorus and 0.053 mug/mL expressed in puerarin for P. lobata. Therapeutic index values is 3.5-20. As a result of in vivo experiments, it was found out that, following therapy using the extracts, animals infected with Acanthamoeba sp. survived for an extended period (2.5-3 times longer). It was determined that plant extracts may be used both externally and internally in the case of a combined therapy for acanthamoebiasis. The tested extracts are not toxic for animals.

PMID: 19050923 [PubMed - as supplied by publisher]

5. Variance of estrogenic activity of the phytoestrogen-rich plant.

Cherdshewasart W, Sriwatcharakul S, Malaivijitnond S.

Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Patumwan, Bangkok 10330, Thailand.

OBJECTIVE: To evaluate the influences of seasonal changes and plant cultivars on estrogenic activity of the phytoestrogen-rich plant, Pueraria mirifica. METHODS: Three cultivars of P. mirifica; PM-I, PM-II and PM-V, were grown in the same field trial for 3 years and random tubers collected during the summer, rainy season and winter seasons. Female Wistar rats were ovariectomized, kept for 14 days, randomly segregated into groups and treated with one of DW, 200mug/100g BW 17beta-estradiol (E2) or tuberous powder of PM-I, PM-II and PM-V at dosages of 100, or 1000mg/kg BW for the next 14 days. For the last 7 days of post-treatment period, rats received only DW. The vaginal cornification was recorded during the treatment and post-treatment period. The uterine tissues of the treated rats at the treatment and post-treatment periods were analyzed for uterine gland number and for the surface area of the myometrium, endometrium and lumen. In addition, ethanol tuberous extracts of PM-I, PM-II and PM-V was submitted to DPPH analysis. RESULTS: Vaginal cornification exhibited a dose-dependent response with plant samples collected during the winter and summer being more active than those collected in the rainy season. All plant samples-induced uterotrophic effects in the analysis at the treatment and post-treatment periods in a dose-dependent manner. The P. mirifica treated rats exhibited increasing uterine gland numbers and thickness of the endometrium and myometrium but a decreasing size of lumen, in comparison to the negative control. The results were more prominent in PM-I than other plants and also in plant samples collected during the winter and summer seasons than in the rainy season. DPPH assay of the ethanol tuberous extracts revealed variance in antioxidant activity. CONCLUSION: The results of uterotrophic and vaginal cornification assays reveal that P. mirifica exhibits a dose-dependent estrogenic activity under the influence of both seasonal changes and plant cultivars, which is confirmed by DPPH assay.

PMID: 18980816 [PubMed - in process]

6. Quantitative aspects of the spread of asian soybean rust in the southeastern United States, 2005 to 2006.

Christiano RS, Scherm H.

ABSTRACT The regional dynamics of soybean rust, caused by Phakopsora pachyrhizi, in six southeastern states (Florida, Georgia, Alabama, South Carolina, North Carolina, and Virginia) in 2005 and 2006 were analyzed based on disease records collected as part of U.S. Department of Agriculture's soybean rust surveillance and monitoring program. The season-long rate of temporal disease progress averaged approximately 0.5 new cases day(1) and was higher in nonsentinel soybean (Glycine max) plots than in sentinel soybean plots and kudzu (Pueraria lobata) plots. Despite the early detection of rust on kudzu in January and/or February each year (representing the final phase of the previous year's epidemic), the disease developed slowly during the spring and early summer on this host species and did not enter its exponential phase until late August, more than 1 month after it did so on soybean. On soybean, cases occurred very sporadically before the beginning of July, after which their number increased rapidly. Thus, while kudzu likely provides the initial inoculum for epidemics on soybean, the rapid increase in disease prevalence on kudzu toward the end of the season appears to be driven by inoculum produced on soybean. Of 112 soybean cases with growth stage data, only one occurred during vegetative crop development while approximately 75% occurred at stage R6 (full seed) or higher. The median nearest-neighbor distance of spread among cases was approximately 70 km in both years, with 10% of the distances each being below approximately 30 km and above approximately 200 km. Considering only the epidemic on soybean, the disease expanded at an average rate of 8.8 and 10.4 km day(1) in 2005 and 2006, respectively. These rates are at the lower range of those reported for the annual spread of tobacco blue mold from the Caribbean Basin through the southeastern United States. Regional spread of soybean rust may be limited by the slow disease progress on kudzu during the first half of the year combined with the short period available for disease establishment on soybean during the vulnerable phase of host reproductive development, although low inoculum availability in 2005 and dry conditions in 2006 also may have reduced epidemic potential.

PMID: 18943512 [PubMed - in process]

9. Decreased expression of aromatase in the Ishikawa and RL95-2 cells by the isoflavone, puerarin, is associated with inhibition of c-jun expression and AP-1 activity.

Li Y, Chen H, Yang S, Xie G.

Cancer Hospital of Tianjin, Tianjin Medical University, Ti-Yuan-Bei, Tianjin 330060, China. liyanwei127@hotmail.com

Aromatase P450 (P450(arom)) is overexpressed in endometriosis, endometrial cancers and uterine fibroids. With weak estrogen agonists/antagonists and some other enzymatic activities, isoflavones are increasingly advocated as a natural alternative to estrogen replacement therapy (ERT) and are available as dietary supplements. Puerarin is major isoflavonoid compound isolated from Pueraria lobata, a Chinese medicine known as Gegen. Our clinical study shows that puerarin can be used in the treatment of endometriosis, which improves pain and infertility. Assuming that the effect of puerarin on endometriosis may result from the regulation of aromatase expression or activity, we carried out this study to test the effects of puerarin on aromatase in Ishikawa and RL95-2 cell lines. Our data have demonstrated a significant decrease of P450(arom) expression at both mRNA and protein levels by low dose puerarin treatment in both cell lines. Besides, we found that the -410/-401bp and -565/-559bp regions of aromatase promoter II contained the critical cis-acting elements, binding AP-1 and c-jun. We also found that puerarin exerted a time-course effect on the inhibition of c-jun mRNA, which parallelled that of P450(arom). To further confirm if c-jun is responsible for the P450(arom) regulation by puerarin, we knocked down c-jun expression using siRNA and it indicates that c-jun acts as a considerable transcription factor in regulating P450(arom) expression and activity. Accordingly, the suppression of P450(arom) expression and activity by puerarin treatment may associate with the downregulation of transcription factor AP-1 or c-jun.

Publication Types: PMID: 18848966 [PubMed - in process]

8. Mechanism of phytoestrogen puerarin-mediated cytoprotection following oxidative injury: estrogen receptor-dependent up-regulation of PI3K/Akt and HO-1.

Hwang YP, Jeong HG.

Department of Pharmacy, College of Pharmacy, Research Center for Proteineous Materials, Chosun University, Gwangju 501-759, Republic of Korea.

Phytoestrogens are polyphenolic non-steroidal plant compounds with estrogen-like biological activity. The phytoestrogen puerarin, the main isoflavone glycoside found in the root of Pueraria lobata, has been used for various medicinal purposes in traditional Chinese medicines for thousands of years. Recent studies have indicated that the estrogen receptor (ER), through interaction with p85, regulates phosphoinositide 3-kinase (PI3K) activity, revealing a physiologic, non-nuclear function of ER that may be relevant in cytoprotection. In this study, we demonstrate that the phytoestrogen puerarin inhibits tert-butyl hydroperoxide (t-BHP)-induced oxidative injury via an ER-dependent Gbeta1/PI3K/Akt and heme oxygenase-1 (HO-1) pathway. Pretreatment of Hepa1c1c7 and HepG2 cells with puerarin significantly reduced t-BHP-induced caspase-3 activation and subsequent cell death. Also, puerarin up-regulated HO-1 expression and this expression conferred cytoprotection against oxidative injury induced by t-BHP. Moreover, puerarin induced Nrf2 nuclear translocation, which is upstream of puerarin-induced HO-1 expression, and PI3K activation, a pathway that is involved in induced Nrf2 nuclear translocation, HO-1 expression and cytoprotection. Puerarin-induced up-regulation of HO-1 and cytoprotection against t-BHP were abolished by silencing Nrf2 expression with specific siRNA. Also, puerarin-mediated increases in PI3K activation and HO-1 induction were reversed by co-treatment with ICI 182,780 and pertussis toxin. Taken together, these results suggest that puerarin augments cellular antioxidant defense capacity through ER-dependent HO-1 induction via the Gbeta1/PI3K/Akt-Nrf2 signaling pathway, thereby protecting cells from oxidative stress.

PMID: 18845176 [PubMed - indexed for MEDLINE]

9. Nootropic activity of tuber extract of Pueraria tuberosa (Roxb).

Rao NV, Pujar B, Nimbal SK, Shantakumar SM, Satyanarayana S.

Department of Pharmacology, V L College of Pharmacy, Raichur 584 103, India. vrngadda@rediffmail.com

Nootropic effect of alcoholic (ALE; 50, 75, 100 mg/kg) and aqueous (AQE; 100, 200, 400 mg/kg) extracts of P. tuberosa was evaluated by using Elevated Plus Maze (EPM), scopolamine-induced amnesia (SIA), diazepam-induced amnesia (DIA), clonidine-induced (NA-mediated) hypothermia (CIH), lithium-induced (5-HT mediated) head twitches (LIH) and haloperidol-induced (DA- mediated) catalepsy (HIC) models. Piracetam was used as the standard drug. A significant increase in inflexion ratio (IR) was recorded in EPM, SIA and DIA models. A significant reversal effect was observed on rectal temperature in CIH model, reduction of head twitches in LIH models. However no significant reduction in catalepsy scores in HIC models were observed with test extracts and standard piracetam. The results indicate that nootropic activity observed with ALE and AQE of tuber extracts of P. tuberosa could be through improved learning and memory either by augmenting the noradrenaline (NA) transmission or by interfering with 5-hydroxytryptamine (5-HT) release. Further, the extracts neither facilitated nor blocked release of the dopamine (DA). Thus ALE and AQE elicited significant nootropic effect in mice and rats by interacting with cholinergic, GABAnergic, adrenergic and serotonergic systems. Phytoconstituents like flavonoids have been reported for their nootropic effect and these are present in both ALE and AQE extracts of tubers of P. tuberosa (Roxb) and these active principles may be responsible for nootropic activity.

PMID: 18814488 [PubMed - in process]

10. Induction of heme oxygenase-1 by plant extract KIOM-79 via Akt pathway and NF-E2 related factor 2 in pancreatic beta-cells.

Kang KA, Kim JS, Zhang R, Piao MJ, Ko DO, Wang ZH, Maeng YH, Eun SY, Hyun JW.

College of Medicine, Cheju National University, Jeju-si, Korea.

The objective of the present study was to determine the mechanism by which KIOM-79 induced heme oxygenase-1 (HO-1) in rat pancreatic beta-cells (RINm5F). A mixture of plant extracts (KIOM-79) was obtained from Magnolia officinalis, Pueraria lobata, Glycyrrhiza uralensis, and Euphorbia pekinensis. HO-1, an antioxidant phase 2 enzyme, was previously reported to possess cytoprotective properties in pancreatic beta-cells. KIOM-79 induced heme oxygenase-1 (HO-1) expression at the mRNA and protein levels, leading to increased HO-1 activity. The transcription factor, NF-E2 related factor 2 (Nrf2), regulates the antioxidant response element (ARE) of the phase 2 detoxifying and antioxidant enzymes, resulting in modulation of HO-1 expression. KIOM-79 increased nuclear translocation, ARE binding, and transcriptional activity of Nrf2. Furthermore, KIOM-79 also elicited activation of Akt (protein kinase B) and LY294004 (inhibitor of Akt)-suppressed KIOM-79-induced activation of Nrf2, which subsequently decreased HO-1 protein levels. Taken together, these data suggest that KIOM-79 augments the cellular antioxidant defense capacity through induction of HO-1 via the Akt-Nrf2-ARE signaling pathway, thereby protecting cells from streptozotocin-induced oxidative stress.

Publication Types: PMID: 18704831 [PubMed - indexed for MEDLINE]

11. Two new isoflavone glycosides from Pueraria lobata.

Sun YG, Wang SS, Feng JT, Xue XY, Liang XM.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.

Two new isoflavone diglycosides, formononetin 8-C-[beta-D-apiofuranosyl-(1 --> 6)]-beta-D-glucopyranoside (1) and formononetin 8-C-[beta-D-xylopyranosyl-(1 --> 6)]-beta-D-glucopyranoside (2), were isolated from the roots of Pueraria lobata, together with four known compounds, 4'-methoxypuerarin (3), daidzin (4), genistin (5), and daidzein (6). The structures of these compounds were elucidated by the spectroscopic methods.

Publication Types: PMID: 18696324 [PubMed - indexed for MEDLINE]

12. Genistein inhibits aldose reductase activity and high glucose-induced TGF-beta2 expression in human lens epithelial cells.

Kim YS, Kim NH, Jung DH, Jang DS, Lee YM, Kim JM, Kim JS.

Department of Herbal Pharmaceutical Development, Korea Institute of Oriental Medicine, 461-24 Jeonmin-dong, Yuseong-gu, Daejeon 305-811, Korea.

Aldose reductase (AR) and TGF-beta have been implicated in the development of diabetic complications, such as cataracts. In an attempt to obtain potential agents for the prevention of diabetic cataracts from natural products, we purified genistein from the roots of Pueraria lobata and investigated its inhibitory effects upon AR activity and its antioxidant effects on rat lenses. The inhibition of AR activity by genistein increased in a dose-dependent manner and the opacities of lenses were significantly improved when treated with genistein. In addition, we determined the effects of genistein on mechanisms induced by exposure to high glucose in human lens epithelial (HLE-B3) cells. We found that genistein was able to reduce the expression of TGF-beta2, alphaB-crystallin, and fibronectin mRNAs in HLE-B3 cells that were cultured in high glucose conditions. In addition, a reduction in glutathione (GSH) levels and thiobarbituratic acid-reactive substances was observed. These results show that genistein is protective against lens opacity and also inhibits high glucose-mediated toxic effects in HLE-B3 cells. These effects are likely achieved by preventing AR and cellular oxidation; therefore, genistein may be a potential therapeutic agent for preventing and treating complications associated with diabetes mellitus, such as diabetic cataracts.

Publication Type PMID: 18692043 [PubMed - indexed for MEDLINE]

13. Isolation of radiolabeled isoflavones from kudzu (Pueraria lobata) root cultures.

Reppert A, Yousef GG, Rogers RB, Lila MA.

Division of Nutritional Sciences, University of Illinois at Urbana-Champaign, 1201 S. Dorner Drive, Urbana, Illinois 61801, USA.

Isoflavones have potential for preventing and treating several chronic health conditions, such as osteoporosis, cardiovascular disease, and cancer. In this study, radiolabeled isoflavones were recovered from kudzu (Pueraria lobata) root cultures after incubation with uniformly labeled (14)C-sucrose in the culture medium for 21 days. Approximately 19% of administered label was recovered in the isoflavone-rich dried extracts of kudzu root cultures (90.2 microCi/g or 3.3 MBq/g extract). HPLC-PDA analysis revealed the predominant isoflavones isolated from kudzu root cultures to be puerarin, daidzin, and malonyl-daidzin. The average concentration of the major isoflavone puerarin in kudzu root cultures was 33.6 mg/g extract, with a specific activity of 63.5 microCi/g (2.3 MBq/g). The isolated isoflavones were sufficiently (14)C-labeled to permit utilization for subsequent in vivo metabolic tracking studies.

Publication Types: PMID: 18690681 [PubMed - indexed for MEDLINE]

14. Puerarin protects PC12 cells against beta-amyloid-induced cell injury.

Zhang HY, Liu YH, Wang HQ, Xu JH, Hu HT.

Department of Human Anatomy and Histology & Embryology, Medical School of Xi'an Jiaotong University, Xi'an, PR China.

beta-Amyloid protein (Abeta), a major protein component of brain senile plaques in Alzheimer's disease, is known to be directly responsible for the production of reactive oxygen species (ROS) and induction of apoptosis. In this study, the protective effect of puerarin, an isoflavone purified from the radix of the Chinese herb Pueraria lobata, on Abeta-induced rat pheochromocytoma (PC12) cultures was investigated. Although exposure of PC12 cells to 50 microM Abeta25-35 caused significant viability loss and apoptotic rate increase, pretreatment of the cells with puerarin for 24h reduced the viability loss and apoptotic rate. Puerarin (1 microM) significantly inhibited Abeta25-35-induced apoptosis of PC12 cells. Preincubation of the cell with puerarin also restored the ROS and mitochondrial membrane potential levels that had been altered as a result of Abeta25-35 treatment. Puerarin was also found to increase the Bcl-2/Bax ratio and reduce caspase-3 activation. These results suggest that puerarin could attenuate Abeta25-35-induced PC12 cell injure and apoptosis and could also promote the survival of PC12 cells. Therefore, puerarin may act as an intracellular ROS scavenger, and its antioxidant properties may protect against Abeta25-35-induced cell injury.

PMID: 18675923 [PubMed - indexed for MEDLINE]

15. One-step rapid determination and purification of puerarin from Radix puerariae by n-octylamine-modified poly(methacrylate-co-ethylene dimethacrylate) monolith.

Lv YQ, Tan TW, Wang MY, Janson JC.

College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, China. yongqinlv.student@sina.com <yongqinlv.student@sina.com>

n-Octylamine-modified poly(methacrylate-co-ethylene dimethacrylate) monoliths were prepared for rapid screening, determination and one-step purification of puerarin from Radix puerariae (a crude extract of the root of Pueraria lobata). The modified monolith showed a specific surface area of 17.8 m(2) g(-1), an average pore size of 0.76 microm and a total porosity of 60.8%. Fast separation of R. puerariae crude extract was achieved within 5 min at a flow velocity of 722 cm h(-1) resulting in a puerarin purity of 97%, with a recovery of 85%. This demonstrates the potential of n-octylamine-modified poly(methacrylate-co-ethylene dimethacrylate) monolith for the rapid analysis and separation of isoflavonoids. Preparative scale sample loading (12 mg in 2 mL) resulted in a purity of 95%, and a recovery of about 69%. HPLC, FTIR, MS and (1)H NMR spectroscopy were used for the characterization and quantification of puerarin in isolated fraction.

Publication Types: PMID: 18639505 [PubMed - indexed for MEDLINE]

16. Estrogenic activity of the dichloromethane extract from Pueraria mirifica.

Sookvanichsilp N, Soonthornchareonnon N, Boonleang C.

Department of Pharmacology, Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand.

Pueraria mirifica and its extracts are widely used as the ingredient(s) in many rejuvenating products. Up to now, the extract of P. mirifica roots that has been used in most studies, is the alcoholic extract. In the present study, we investigated the estrogenic activity using uterotropic and MCF-7 cell proliferation models of the dichloromethane extract as well as the water extract which was obtained from partitioning the ethanolic extract. The results indicated that among the three extracts, i.e. the ethanolic extract (PM1), the water extract (PM2) and dichloromethane extract (PM3), PM3 exhibited the most potent estrogenic activity in both models, followed by PM1. The extracts produced uterotropic activity associated with the increase of water content while uterotropic activity of 17beta-estradiol was related to the increase of muscle mass. The two isoflavonoids, genistein and daidzein, were not the major active phytoestrogens involving the estrogenic activity of these extracts.

PMID: 18621111 [PubMed - in process]

17. Batch foam fractionation of kudzu (Pueraria lobata) vine retting solution.

Eiamwat J, Loha V, Prokop A, Tanner RD.

Department of Chemical Engineering, Vanderbilt University, 37235, Nashville, TN.

The aqueous protein solution from kudzu(Pueraria lobata) vine retting broth, without the addition of other surfactants, was foam-fractionated in a vertical tubular column with multiple sampling ports. Time-varying trajectories of the total protein levels were determined to describe the protein behavior at six positions along the 1-m column. The lowest two trajectories of this batch process represented a loss of proteins from the bulk liquid and tended to merge and decay together in time; the other trajectories displayed a gain in proteins in the foam phase. These upper column port protein concentration trajectories generally increased in time up to 45 min, followed by a decrease, reflecting the removal of proteins from the column ports. The foam became dryer as it passed up the column to the top port. The protein concentration was about 5-8xhigher in the top port foam than in the initial bulk solution, mainly as a result of liquid drainage from the foam along the column axis. This concentration increase in the collected foam was dependent on the initial pH of the bulk solution. The mol-wt profile of the proteins in the concentrated foam effluent was determined by one-dimensional gel electrophoresis. An analysis of the gel electropherograms indicated that the most abundant proteins could be cellulases and pectinases.

PMID: 18576021 [PubMed - in process]

18. Safety of health foods and importance of their origin.

Goda Y.

National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, Japan. goda@nihs.go.jp

The safety guideline for voluntary inspections on the ingredients used for capsulated or pellet food, announced by the director of the department of food safety of the Ministry of Health, Labor and Welfare on February 1, 2008 states that "how to guarantee the origin" is the top priority to ensure safety. However, in the course of our continuous investigation of the origin of natural products, the ingredients of some health food products such as chondroitin sulfate, white kwao keur (Pueraria candollei var. mirifica) and black cohosh did not originate from the labeled material. The usage of the correct origin is the fist step for the quality assurance of "health food". Therefore, we believe that regulatory requirements for accurately indicating the origin of "health foods" and effective enforcement of these requirements are needed.

Publication Types: PMID: 18520130 [PubMed - indexed for MEDLINE]

19. Effects of botanical dietary supplements on cardiovascular, cognitive, and metabolic function in males and females.

Carlson S, Peng N, Prasain JK, Wyss JM.

Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama, USA.

BACKGROUND: The onset of menopause marks a pivotal time in which the incidence of hypertension and of cardiovascular disease (CVD) begins to increase dramatically in women. Before menopause, the incidences of these diseases are significantly lower in women than in age-matched men. After menopause, the rates of these diseases in women eventually approximate those in men. The loss of endogenous estrogen at menopause has been traditionally believed to be the primary factor involved in these changes. OBJECTIVE: This review summarizes recent findings regarding the effectiveness of botanicals in the treatment of some menopausal symptoms and other symptoms of aging (eg, rise in arterial pressure, cognitive decline, insulin resistance, and hyperlipidemia). METHODS: Articles were selected for inclusion in this review based on the significance of the research and contribution to the current understanding of how each botanical elicits cardioprotective effects. To this end, PubMed and MEDLINE databases were searched, using terms that included the name of the specific botanical along with the relevant aspects of its action(s), such as blood pressure, glycemic control, and lipids. Most of the articles used were published within the past 5 years, although some older articles that were seminal in advancing the current understanding of botanicals were also included. RESULTS: Soy has been found to lower plasma lipid concentrations and arterial pressure in postmenopausal women and age-matched men, and to have protective effects in heart disease and atherosclerosis of the carotid and coronary circulation. Soy was also found to lower fasting insulin concentrations and glycosylated hemoglobin concentrations. Grape seed extract, another frequently used botanical, contains polyphenols that have been found to reduce arterial pressure and salt-sensitive hypertension in estrogendepleted animal models. CONCLUSION: Several botanical compounds have been found to have beneficial effects in the treatment of the symptoms of menopause and other symptoms of aging, including CVD, cognitive decline, and metabolic diseases.

Publication Types: PMID: 18395685 [PubMed - indexed for MEDLINE]

20. Simultaneous determination of ten active components in traditional Chinese medicinal products containing both Gegen (Pueraria lobata) and Danshen (Salvia miltiorrhiza) by high-performance liquid chromatography.

Chang Q, Sun L, Zhao RH, Chow MS, Zuo Z.

Institute of Medicinal Plant Development, Chinese Academy of Medical Science & Peking Union Medical College, Beijing, 100094, People's Republic of China.

In order to facilitate the quality control of Traditional Chinese Medicine (TCM) products containing both Gegen (Pueraria lobata) and Danshen (Salvia miltiorrhiza), a new and simple HPLC method for the simultaneous determination of 10 active components in these products has been developed. The chromatographic separation was carried out on a C(18) column eluted with a mobile phase consisting of 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile with gradient elution. The eluent was monitored by a photodiode array UV detector at a wavelength of 250 nm for Gegen components including puerarin, daidzein 8-C-apiosyl-glucoside, daidzin and daidzein, and at 270 nm for Danshen components including danshensu, protocatechuic aldehyde, salvianolic acid B, cryptotanshinone, tanshinone I and tanshinone IIa. Excellent chromatographic separation was achieved for all studied compounds with good linearity (r(2)> 0.999) over the studied concentration ranges. The developed method has been applied to the simultaneous determination of the 10 studied compounds in commercially available products containing both Gegen and Danshen. The TCM product samples were extracted by sonication with a mixture of methanol:water (80:20) containing 0.5% acetic acid. Extraction recoveries for all studied compounds were in the range of 96.01-106.18%. The intra-day and inter-day variations were less than 7.25 and 5.44%, respectively, for all studied compounds. The developed method has not only proved to be effective in the simultaneous determination of the 10 components, but also provides a convenient quality control approach for TCM products containing both Gegen and Danshen.

Publication Types: PMID: 18438756 [PubMed - indexed for MEDLINE]

21. HILIC for separation of co-eluted flavonoids under RP-HPLC mode.

Zhang H, Guo Z, Zhang F, Xu Q, Liang X.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences Dalian, China.

Hydrophilic interaction chromatography (HILIC) was employed to separate the co-eluted flavonoids from licorice extract under RP-HPLC mode. HILIC separations were carried out with the Atalantis HILIC Silica column and the CD-based column. The co-eluted flavonoids were well retained and separated on the two HILIC columns under HILIC mode. Similar results were obtained in the separation of another isoflavones sample, from kudzu extract under HILIC mode.

Publication Types: PMID: 18428179 [PubMed - indexed for MEDLINE]

22. Pueraria mirifica, phytoestrogen-induced change in synaptophysin expression via estrogen receptor in rat hippocampal neuron.

Chindewa R, Lapanantasin S, Sanvarinda Y, Chongthammakun S.

Center for Neuroscience, Faculty of Science, Mahidol University, Bangkok, Thailand.

OBJECTIVE: To examine Pueraria mirifica (Leguminosae) containing-phytoestrogen effect on synaptic density and involvement of estrogen receptor. MATERIAL AND METHOD: The level of synaptophysin, a presynaptic vesicle protein, was measured using Western blot analysis and immunocytochemistry in hippocampal primary cell cultures at 6 days in vitro. RESULTS: P. mirifica and 17beta-estradiol (0.1 microM) treatment for 4 days, but not for 2 days, significantly increased synaptophysin immunoreactivity and level of synaptophysin. P. mirifica up to 60 microg/ml resulted in a dose related increase in the level of synaptophysin immunoreactivity. The classical estrogen receptor antagonist, ICI 182 780, significantly blocked P. mirifica-induced increase in synaptophysin. CONCLUSION: P. mirifica-containing phytoestrogen affects synaptic density by inducing synaptophysin expression via estrogen receptor.

Publication Types: PMID: 18389986 [PubMed - indexed for MEDLINE]

23. Effect of multiple mating on reproduction and longevity of the phytoseiid mite Neoseiulus californicus.

Gotoh T, Tsuchiya A.

Laboratory of Applied Entomology and Zoology, Faculty of Agriculture, Ibaraki University, Ami, Ibaraki 300-0393, Japan. gotoh@mx.ibaraki.ac.jp

Females of some insects and mites that mate repeatedly (multiple-mated females) or that have limited food supply have a shortened life span, compared to females that are allowed to mate only once (single-mated females) or that have ample food. The present study focused on the relationship between mating frequency and various reproduction-related parameters in a commercial strain (Spical) of the phytoseiid mite Neoseiulus californicus (McGregor) (Acari: Phytoseiidae). The impetus for this study came from the previous finding that single-mated females displayed an extremely long postoviposition period (ca. 60 days), accounting for more than 75% of the total adult longevity. In the present study it was found that the postoviposition period in the multiple-mated females was shorter than in the single-mated females and that longevity was gradually shortened with increasing mating frequency. Total egg production in females that mated twice or more was higher than in single-mated females, but egg production did not increase further with more than two matings. Single- and multiple-mated females ceased oviposition on the 24th and 34th day of adult life, respectively; multiple-mated females produced only male progeny on and after the 30th day. The postoviposition period of multiple-mated females was 40% shorter than that of single-mated females. Yet, the postoviposition period of the multiple-mated females is still more than 30 days, suggesting that other factors besides mating frequency have an influence on the prolonged postoviposition period. The reproductive ability of the Spical strain females gradually decreased with increasing virgin period (i.e. with increasing age), but they did not lose their reproductive potential--not even when mating was delayed for more than 2 months (i.e. females remained virgin for 70 days).

PMID: 18379888 [PubMed - indexed for MEDLINE]

24. Metabolic activation promotes estrogenic activity of the phytoestrogen-rich plant.

Cherdshewasart W, Sriwatcharakul S.

Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Patumwan, Bangkok 10330, Thailand. cwichai@sc.chula.ac.th

OBJECTIVE: There is evidence that metabolic activation can increase the estrogenic activity of the phytoestrogen-rich herb in tests with HepG2 cells. Variation in both plant genetics and harvest season may also influence estrogenic activity of the plant materials. We evaluated the influence of in vitro metabolic activation by S9 mixture on the estrogenic activity of tuberous samples of different cultivars of the phytoestrogen-rich herb, Pueraria mirifica, harvested in different seasons. METHODS: Plant extracts were derived from the tubers of five plant cultivars collected during summer, rainy season and winter and administered to MCF-7 cultures, an ERalpha-positive human mammary adenocarcinoma cell line for 3 days at dosages of 0.1, 1, 10, 100 and 1000microg/ml. These data were compared with the major plant isoflavonoids puerarin, daidzin, genistin, daidzein and genistein and with 17beta-estradiol, at concentrations of 10(-12) to 10(-6)M. The test system was done in the absence and presence of the S9 mixture. RESULTS: The major plant isoflavonoids and the plant extracts exhibited variable degrees of estrogenic activities as evaluated by altered proliferation of the MCF-7 cell line which were significantly enhanced in the presence of the S9 mixture. CONCLUSION: Metabolic activation of plant isoflavonoids at least in vitro by S9 mixture plays a significant role in amplification of the estrogenic activity of the phytoestrogen-rich plant. In addition, the estrogenic activities of the plant samples were potentially influenced by both seasonal changes and plant genetics.

Publication Types: PMID: 18313242 [PubMed - indexed for MEDLINE]

25. Preventive effects of Pueraria mirifica on bone loss in ovariectomized rats.

Urasopon N, Hamada Y, Cherdshewasart W, Malaivijitnond S.

Biological Science Ph.D. Program, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.

OBJECTIVE: Effects of Pueraria mirifica on bone loss in fully mature ovariectomized rats are examined. METHODS: Two series of experiments were performed. In the first series, rats were kept with their ovaries intact and divided into two groups; initial control (IC) and sham control (SH). The IC rats were sacrificed on day 1 and their data were kept as baseline control. The SH rats were subjected to sham operation on day 0 and gavaged daily with distilled water for 90 days. In the second series, rats were subjected to ovariectomy, divided into five groups and gavaged daily with 0.1mg/kg B.W./day of 17-alpha-ethinylestradiol (EE), 0, 10, 100 and 1000mg/kg B.W./day of P. mirifica (P0, P10, P100 and P1000, respectively) for 90 days. Changes of bone mineral density and bone mineral content were measured using peripheral Quantitative Computerized Tomography. RESULTS: Bone loss was significantly induced by ovariectomy and it was dose-dependently prevented by P. mirifica treatment for 90 days. The preventive effects of P. mirifica on bone loss depended on bone types (axial or long bone), bone sites (metaphysis or diaphysis), and bone compartments (trabecular and cortical). At P100 and P1000, bone loss was completely prevented both in trabecular bone mineral density and content. The effects of P. mirifica were, as expected, comparable to that in the EE group. CONCLUSION: These results suggest that P. mirifica may be applicable to treat the osteoporosis in menopausal women; however, an undesirable side effect on stimulating reproductive organs should be concerned.

Publication Types: PMID: 18313241 [PubMed - indexed for MEDLINE]

26. Biology and preliminary host range assessment of two potential kudzu biological control agents.

Frye MJ, Hough-Goldstein J, Sun JH.

Department of Entomology and Wildlife Ecology, University of Delaware, Newark, DE 19716-2160, USA.

Two insect species from China, Gonioctena tredecimmaculata (Jacoby) (Coleoptera: Chrysomelidae) and Ornatalcides (Mesalcidodes) trifidus (Pascoe) (Coleoptera: Curculionidae), were studied in quarantine in the United States as potential biological control agents for kudzu, Pueraria montana variety lobata (Willd.) Maesen and S. Almeida. Adults of G. tredecimmaculata were ovoviviparous and reproduced throughout the summer, producing offspring that had an obligate adult diapause. In no-choice tests, adult and larval G. tredecimmaculata rejected most of the plant species tested, but consumed foliage and completed their life cycle on soybean (Glycine max L. Merr.) and on a native woodland plant, hog-peanut (Amphicarpaea bracteata L. Fernald), which are in the same subtribe as kudzu (Glycininae). Insects showed similar responses to field- and greenhouse-grown soybean and kudzu foliage, despite measurable differences in leaf traits: field-grown foliage of both plants had greater leaf toughness, higher total carbon content, higher trichome density, and lower water content than greenhouse foliage. O. trifidus adults also rejected most of the plants tested but fed on and severely damaged potted soybean and hog-peanut plants in addition to kudzu. Further tests in China are needed to determine whether these species will accept nontarget host plants under open-field conditions.

Publication Types: PMID: 18284771 [PubMed - indexed for MEDLINE]

27. [Pharmacokinetics of pueraria flavonoids from Pueraria lobata by anti-oxidant pharmacodynamics effect method]

[Article in Chinese]

Jia XB, Cai Y, Chen Y, Cheng XD, Fan CY, Shi YF.

Jiangsu Academy of Chinese Medicine Sciences, Jiangsu Engineering and Technology Research Center for Modern Chinese Pharmaceutical Preparation, Nanjing 210028, China.

OBJECTIVE: To study the pharmacokinetics of pueraria flavonoids(PF) from Pueraria lobata. METHOD: An anti-oxidant pharmacodynamics effect method was quoted to detect the antioxidant ability of PF contained in rats plasm after oral delivered, measures the fluorescence intensity, and calculates the concentration of components in plasma, then we can get the plot of the change of the content of PF contained in rats plasm, and then we can get the pharmacokinetics of Pueraria Flavonoids pellets. RESULT: The AUC of Pueraria lobata isoflavone pellets is 4.40-fold of Yufengningxin tablets. CONCLUSION: It can enhance the bioavailability of PF by made it to pellets.

Publication Types: PMID: 18257261 [PubMed - in process]

28. Effect of chemical factors on production of isoflavonoids in Pueraria tuberosa (Roxb.ex.Willd.) DC suspension culture.

Goyal S, Ramawat KG.

Laboratory of Biomolecular Technology, Department of Botany, M.L. Sukhadia University, Udaipur 313 001, India.

Suspension cultures of Pueraria tuberosa, a woody legume, have been established and using different concentrations of growth regulators, sucrose, ammonium and nitrate nitrogen, attempts have been made to improve their isoflavonoid content. The cell cultures grew well on all the treatments. Up to approximately 8 folds increased isoflavonoids content was recorded in the cultures grown in MS medium modified with nitrogen and supplemented with 1 mg 1(-1) of kinetin.

Publication Types: PMID: 18254213 [PubMed - indexed for MEDLINE]

29. [Pharmacokinetics of pueraria for intranasal on spray in rabbits]

[Article in Chinese]

Lu Y, Zhang T, Tao JS, Xu LY.

Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.

OBJECTIVE: To explore pharmacokinetic features of puerarin in pueraria spray and calculate pharmacokinetic parameters according to puerarin of drug-time curve in rabbits. METHODS: The concentration of puerarin in plamsa was determined by HPLC. The methanol was used to sediment protine. The 3P87 program was used to calculate the pharmacokinetic parameters. RESULTS: The vivo course of pueraria in spray could be described by the two compartment model. The main pharmacokinetic parameters of Pueraria spray were: t(l/2(beta)) =0.93 h, CL =44.23 mg x L(-1), AUC = 16.28 mg x h x L(-1), Cmax =5.9 mg x L(-1) and tmax = 0.975 h. CONCLUSION: The study will provide some scientific basises for the quality evaluation and pharmaceutics reformation of pueraria for intranasal.

Publication Types: PMID: 18236755 [PubMed - indexed for MEDLINE]

30. Effects and safety of Pueraria mirifica on lipid profiles and biochemical markers of bone turnover rates in healthy postmenopausal women.

Manonai J, Chittacharoen A, Udomsubpayakul U, Theppisai H, Theppisai U.

Department of Obstetrics and Gynaecology, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand. rajmo@mahidol.ac.th

OBJECTIVE: To evaluate the effect of Pueraria mirifica on lipid profiles and biochemical markers of bone turnover rates in healthy postmenopausal women and to evaluate the safety of Pueraria mirifica on endometrium; breast tissue; and hematologic, hepatic, and renal systems. DESIGN: This was a randomized, double-blind, placebo-controlled study in a university hospital of healthy postmenopausal women aged 45 to 60 years old. Women were enrolled voluntarily and randomly received 20, 30, or 50 mg Pueraria mirifica in capsules or identical placebo once daily for 24 weeks. Outcome measures were lipid profiles, bone-specific alkaline phosphatase level, endometrial thickness, endometrial histology, breast ultrasonography, complete blood count, liver function test, and renal function test. RESULTS: After 24 weeks of treatment, 71 women were evaluated. Of the 71 women, 51 randomly received varying doses of Pueraria mirifica and 20 received placebo. Pueraria mirifica and placebo significantly increased triglyceride levels by 15% from baseline levels (P<0.05). The Pueraria mirifica group showed a significant decrease in bone-specific alkaline phosphatase levels after 24 weeks of treatment compared with the placebo group; from 0.22+/-0.18 U/L to 0.13+/-0.01 U/L in the Pueraria mirifica group and from 0.20+/-0.10 U/L to 0.20+/-0.14 U/L in the placebo group. Endometrial thickness did not change after treatment in both groups (P>0.05). No endometrial proliferation or hyperplasia was reported after 24 weeks of treatment in both groups. There were no significant differences in adverse effects on breast tissue, complete blood count, and liver and renal function tests between the Pueraria mirifica and placebo groups in this study. CONCLUSION: Pueraria mirifica at a dose of 20, 30, and 50 mg/d for a 24-week period demonstrated an estrogen-like effect on bone turnover rate. Pueraria mirifica did not demonstrate an estrogen-like effect on endometrial thickness and endometrial histology. Mild adverse effects occurred after Pueraria mirifica and placebo treatment.

Publication Types: PMID: 18202589 [PubMed - indexed for MEDLINE]

31. Determination of the estrogenic activity of wild phytoestrogen-rich Pueraria mirifica by MCF-7 proliferation assay.

Cherdshewasart W, Traisup V, Picha P.

Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand. cwichai@sc.chula.ac.th

The aim of this study was to evaluate the estrogenic activity of tuberous samples of wild, phytoestrogen-rich Pueraria mirifica collected from 28 out of 76 provinces of Thailand by MCF-7 proliferation assay. The plant extracts were administered to MCF-7, ER alpha positive human mammary adenocarcinoma cell cultures, for 3 days at dosages of 0.1, 1, 10, 100 and 1,000 microg/ml and were compared with 17 beta-estradiol at concentrations of 10(-12)-10(-6) M. The mean P. mirifica population at 1 mug/ml exhibited significant proliferation. Two plant samples exhibited levels of proliferation in MCF-7 that were similar to 17beta-estradiol. The mean P. mirifica populations at 100 and 1,000 microg/ml exhibited significant cytotoxicity in MCF-7. Analysis of the estrogenic activity of puerarin, representative of major isoflavonoids in P. mirifica tubers, revealed proliferation in MCF-7 only at the highest dose (10(-6) M) that was 10(2)-10(5) times less active than 17 beta-estradiol. Puerarin and 17 beta-estradiol at concentration of 10(-12)-10(-6) M exhibited no cytotoxicity in MCF-7.

Publication Types: PMID: 18160771 [PubMed - indexed for MEDLINE]

32. [Study on cryopreservation of Pueraria lobata germplasm by vitrification]

[Article in Chinese]

Hong SR, Yin MH.

Life Sciences Department, Shangrao Normal College, Shangrao 334001, China. hongsenrong@163.com

The experiment of the cryopreservation technique on Pueraria lobata showed that: At first, the aseptic explants were treated at 4 degrees C for 5 days. Then the stems with buds were cut and precultured at 4 degrees C for 1 day in 5% DMSO + 5% sucrose media. They were dehydrated with 60% and 100% PVS2 (30% glycerol + 15% glycol + 15% dimethyl sulfoxide + 0.4 mol/L sucrose) at 0 degrees C for 30 minutes respectively. At last the stems were immersed immediately into liquid nitrogen directly and conserved for 24 hours. After rapidly thawing in a water bath at 40 degrees C for 90 seconds, the stems were washed two times with MS media supplemented with 1.2 mol/L sucrose and 10 minutes each time, then transferred on the MS media supplemented with BA 2 mg/L and NAA 1 mg/l, in dark for 7 days and then in light. The survival rate was up to 57-58% . The regenerated plant of Pueraria lobata were the same as the normal in morphology.

Publication Types: PMID: 18074831 [PubMed - indexed for MEDLINE]

33. Isolation and characterization of nonrhizobial plant growth promoting bacteria from nodules of Kudzu (Pueraria thunbergiana) and their effect on wheat seedling growth.

Selvakumar G, Kundu S, Gupta AD, Shouche YS, Gupta HS.

Vivekananda Institute of Hill Agriculture, Indian Council of Agricultural Research, Almora 263601, Uttarakhand, India. gselva74@rediffmail.com

The leguminous vine Kudzu (Pueraria thunbergiana) is an introduction into the N. W. Himalayan region of India. Despite its value as a fodder and cover crop, little is known about the nature of the nodule microflora. In an attempt to study the nodule bacteria, we isolated and characterized three nonrhizobial plant growth promoting bacteria from surface sterilized nodules of Kudzu. Based on the sequencing of the 16 S r RNA gene, the isolates were designated as Bacillus thuringiensis KR-1, Enterobacter asburiae KR-3, and Serratia marcescens KR-4. Crystalline bodies were detected in the isolate KR-1, confirming its identity as B. thuringiensis. Under in vitro conditions, all three isolates were found to produce indole acetic acid. Other plant growth promotion attributes such as P solubilization, hydrogen cyanide production, and ammonia production varied among the isolates. All of the three isolates promoted growth and positively influenced nutrient uptake parameters of wheat seedlings.

PMID: 18074178 [PubMed - indexed for MEDLINE]

34. Development and validation of an HPLC method for quality control of Pueraria lobata flower.

Bebrevska L, Bravo L, Vandervoort J, Pieters L, Vlietinck A, Apers S.

Laboratory of Pharmacognosy and Pharmaceutical Analysis, Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium. Lidiya.bebrevska@ua.ac.be

Pueraria lobata, also known as Kudzu (Japan) or Ge (China), is a medicinal plant widely used in Oriental traditional medicine. In this study the development, optimization and validation of an HPLC ethod for quality control of Pueraria flower plant material is presented. By means of this analytical method the three major compounds, i. e., the isoflavones tectorigenin 7- O-[beta- D-xylopyranosyl-(1 - 6)-beta- D-glucopyranoside], tectorigenin 7- O-beta- D-glucopyranoside and tectorigenin, were quantified, using the isoflavones genistin and genistein as external standards. The extraction procedure, the extraction solvent, the extraction yields and the HPLC conditions were evaluated and optimized. The samples were analyzed on an RP C18 column, and eluted with a binary system consisting of water and methanol using a linear gradient; detection was at 262 nm. Tectorigenin used in the recovery experiments was isolated and purified in the laboratory. The final method was fully validated according to the ICH guidelines in terms of linearity, precision and accuracy. The validation data showed that the precision, (RSD% (betweendays) of 3.1, 2.84 and 1.77 for the three major compounds, respectively), and the accuracy (recovery of 104.2 %) were acceptable. These validation results demonstrate the suitability of the method for the quality control of this crude drug.

Publication Types: PMID: 18067064 [PubMed - indexed for MEDLINE]

35. [A new chemical component of Pueraria lobata (Willd.) Ohwi]

[Article in Chinese]

Wang YZ, Feng WS, Shi RB, Liu B.

School of Pharmacy, Henan College of Traditional Chinese Medicine, Zhengzhou 450008, China. wangyzlb@126.com

Pueraria lobata (Willd.) Ohwi. was extracted for two times with 70% ethanol and the 70% ethanol-extracts was condensed. Various column chromatography with AB-8 macroreticular resin, Toyopearl HW-40, pharmadex LH-20, and silica gel were employed for the isolation and purification of the 70% ethanol-extracts from Pueraria lobata (Willd.) Ohwi. Five compounds were isolated and their structures were identified by physiochemical properties and spectral analysis (UV, IR, MS, 1H NMR, 13C NMR, HMQC, HMBC, etc.): (4R)-3-[ 2-hydroxy-4-methoxyphenyl]-4-(4-beta-D-glucopyranosyloxybenzyl) but-2-en-4-olide (1), 4', 8-dimethoxyl-7-O-beta-D-glucopyranosyl isoflavone (2), eicosanoic acid (3), hexadecanoic acid (4), tetracosanoid acid-2,3-dihydroxypropyl ester (5). Compound 1 was a new compound, and compounds 2, 3, 4 were isolated from this plant for the first time.

Publication TypesPMID: 18050739 [PubMed - in process]

36. Protective effects of puerarin on carbon tetrachloride-induced hepatotoxicity.

Hwang YP, Choi CY, Chung YC, Jeon SS, Jeong HG.

Department of Pharmacy, College of Pharmacy, Research Center for Proteineous Materials, Chosun University, Kwangju, Korea.

Puerarin, the main isoflavone glycoside found in the root of Pueraria lobata, has been used for various medicinal purposes in traditional Chinese medicine for thousands of years. The purpose of this study was to investigate the protective effects of puerarin against hepatotoxicity induced by carbon tetrachloride (CCl4) and the mechanism of its hepatoprotective effect. In mice, pretreatment with puerarin prior to the administration of CCl4 significantly prevented the increased serum enzymatic activity of alanine aspartate aminotransferase and hepatic malondialdehyde formation in a dose-dependent manner. In addition, pretreatment with puerarin significantly prevented both the depletion of reduced glutathione (GSH) content and the decrease in glutathione S-transferase (GST) activity in the liver of CCl4-intoxicated mice. Hepatic GSH levels and GST activity were increased by treatment with puerarin alone. CCl4-induced hepatotoxicity was also prevented, as indicated by liver histopathology. The effects of puerarin on cytochrome P450 (CYP) 2E1, the major isozyme involved in CCl4 bioactivation, were also investigated. Treatment of the mice with puerarin resulted in a significant decrease in the CYP2E1-dependent aniline hydroxylation in a dose-dependent manner. Consistent with these observations, the CYP2E1 protein levels were also lowered. Puerarin exhibited anti-oxidant effects on FeCl2-ascorbate induced lipid peroxidation in mouse liver homogenates, and on superoxide radical scavenging activity. These results suggest that the protective effects of puerarin against the CCl4-induced hepatotoxicity possibly involve mechanisms related to its ability to block CYP-mediated CCl4 bioactivation, induction of GST activity and free radical scavenging effects.

Publication Types: PMID: 18038910 [PubMed - indexed for MEDLINE]

37. How many symbionts are provided by mothers, acquired by offspring, and needed for successful vertical transmission in an obligate insect-bacterium mutualism?

Hosokawa T, Kikuchi Y, Fukatsu T.

Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan. th-hosokawa@aist.go.jp

Vertical symbiont transmission is among the most pivotal processes for maintenance of symbiotic associations. However, it is poorly understood whether and how the levels of resource allocation and investment upon vertical transmission are regulated. The stinkbug Megacopta punctatissima is obligatorily associated with the gut symbiotic bacterium 'Candidatus Ishikawaella capsulata', whose transmission is mediated by a unique mechanism called 'symbiont capsule'. We investigated the population dynamics of the symbiont during vertical transmission in the host-symbiont mutualism. The stinkbug mothers produced one capsule for around 3.6 eggs irrespective of clutch size, suggesting a strict maternal control over symbiont supply for the offspring. However, experimental manipulation of egg/capsule ratios revealed that one capsule is sufficient for symbiont transmission to six nymphs. Quantitative polymerase chain reaction analyses demonstrated that a capsule contains 1.2 x 10(8) symbionts, a newborn nymph possesses 2 x 10(7) symbionts from a capsule, and thus one capsule certainly contains a sufficient amount of symbiont cells for six nymphs. These results indicated that the stinkbug mothers produce 1.7 times more symbiont capsules than needed. The newborn nymphs consistently harboured around 2 x 10(7) symbionts, also suggesting a nymphal control over symbiont transmission. The threshold symbiont titre minimally needed for successful vertical transmission was estimated to be 1.9 x 10(6) symbionts, which is only 1/10 of the actual symbiont titre detected in a newborn nymph. These results illuminate several ecological factors that may be relevant to parental and offspring controls over symbiotic resource allocation through host insect generations.

Publication Types: PMID: 18028305 [PubMed - indexed for MEDLINE]

38. [Study of HPLC-DAD fingerprint on complex traditional Chinese medicine proprietary preparation-Baoji pills]

[Article in Chinese]

Wang XH, Xie PS, Tian RT, Huang XD, Zheng RB, Qin CM, Yu QX.

Zhuhai Chromap Institute of Herbal Medicine Research, Zhuhai 519085, China.

OBJECTIVE: Based on 'Back-tracking' method, identification and quality evaluation of complex traditional Chinese medicine (TCM) preparation of Baoji pills (BJP) were carried out by HPLC fingerprint analysis. METHOD: HPLC-DAD fingerprint of BJP was conducted with Zorbax SB-C18 column and non-linear elution with the mobile phase consisted of acetonitrile-0.5% glacial acetic acid at column temperature 30 degrees C and detective wavelengths of 250 nm and 283 nm. From the established chromatographic pattern of BJP, track backward to the corresponding crude herbal drugs in the formula, attribution ofmost peaks in the BJP fingerprint can be disclosed. RESULT: The BJP HPLC fingerprint consisted of 44 peaks among which 35 peaks were assigned by parallel comparison with the fingerprint of the 10 corresponding crude drugs in the formula such as pueraria, pummelo peel, and magnolia bark, etc. and 22 peaks we reidentified by comparison with the chemical reference substances. CONCLUSION: The established HPLC fingerprint represents the whole character of BJP, which enhanced the specialty for control and assessment of the product quality. It exemplified much more effective for quality control than selecting any marker for qualitative or quantitative testing target. And the Back-tracking' experimental method extended the study mentality for complex formula TCM products chromatographic fingerprinting analysis.

Publication Types: PMID: 17992991 [PubMed - indexed for MEDLINE]

39. Pueraria lobata (Kudzu root) hangover remedies and acetaldehyde-associated neoplasm risk.

McGregor NR.

University of Melbourne, Faculty of Medicine, Dentistry and Health Sciences, 9 Auburn Grove, Armadale, Victoria 3143, Australia. NeilM@unimelb.edu.au

Recent introduction of several commercial Kudzu root (Pueraria lobata) containing hangover remedies has occurred in western countries. The available data is reviewed to assess if there are any potential concerns in relationship to the development of neoplasm if these products are used chronically. The herb Pueraria has two components that are used as traditional therapies; Pueraria lobata, the root based herb and Pueraria flos, the flower based herb. Both of these herbal components have different traditional claims and constituents. Pueraria flos, which enhances acetaldehyde removal, is the traditional hangover remedy. Conversely, Pueraria lobata is a known inhibitor of mitochondrial aldehyde dehydrogenase (ALDH2) and increases acetaldehyde. Pueraria lobata is being investigated for use as an aversion therapy for alcoholics due to these characteristics. Pueraria lobata is not a traditional hangover therapy yet has been accepted as the registered active component in many of these hangover products. The risk of development of acetaldehyde pathology, including neoplasms, is associated with genetic polymorphism with enhanced alcohol dehydrogenase (ADH) or reduced ALDH activity leading to increased acetaldehyde levels in the tissues. The chronic usage of Pueraria lobata at times of high ethanol consumption, such as in hangover remedies, may predispose subjects to an increased risk of acetaldehyde-related neoplasm and pathology. The guidelines for Disulfiram, an ALDH2 inhibitor, provide a set of guidelines for use with the herb Pueraria lobata. Pueraria lobata appears to be an inappropriate herb for use in herbal hangover remedies as it is an inhibitor of ALDH2. The recommendations for its use should be similar to those for the ALDH2 inhibitor, Disulfiram.

Publication Types: PMID: 17980785 [PubMed - indexed for MEDLINE]

40. Efficacy comparison of Pueraria mirifica (PM) against conjugated equine estrogen (CEE) with/without medroxyprogesterone acetate (MPA) in the treatment of climacteric symptoms in perimenopausal women: phase III study.

Chandeying V, Sangthawan M.

Department of Obstetrics and Gynecology, Faculty of Medicine, Prince of Songkhla University, Hat Yai, Songkla 90112, Thailand. verapol.c@psu.ac.th

OBJECTIVE: To evaluate the efficacy comparison of Pueraria mirifica (PM), name in Thai is Kwao Kruea Khao, against conjugated equine estrogen (CEE) with/without medroxyprogesterone acetate (MPA) in the treatment of perimenopuasal women with climacteric symptoms. MATERIAL AND METHOD: Perimenopausal women attending the Menopausal clinic of Hat Yai Regional Hospital were voluntarily recruited. The vasomotor symptoms such as hot flushes and night sweats, as well as other unpleasant symptoms, urogenital and psychological symptoms, were also assessed. Patients were voluntarily enrolled and randomly received daily 50 mg raw material of PM, Group A, or daily 0.625 mg of conjugated equine estrogen (CEE) with/without 2.5 mg of medroxyprogesterone acetate (MPA), Group B, depend on non-hysterectomized/hysterectomized condition. RESULTS: Seventy-one patients were enrolled. Eleven of those were excluded for failing to complete the initial work-up and follow-up. Sixty cases were evaluated, 30 cases in Group A and 30 cases in Group B. After medication, the mean of modified Greene climacteric scale (MGCS) in Group A/Group B had decreased from 29.0/32.26 to 17.86/18.1, 12.56/9.57 and 9.9/8.16 at 1-, 3-, and 6- month respectively. The clinical satisfaction using MGCS was not statistically significant between PM (Group A) and CEE with/without MPA (Group B) in the alleviation of climacteric symptoms (p-value > 0.05). There were no statistically significant changes of three serum markers: estradiol, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) between both groups. CONCLUSION: PM, containing phytoestrogens, has estrogenic effect as similar as CEE, and can alleviate the climacteric symptoms in perimenopausal women. PM demonstrates great promise in the treatment of climacteric symptoms. However, optimal doses should be clinically assessed to meet appropriate individual responses.

Publication Types: PMID: 17957910 [PubMed - indexed for MEDLINE]

41. [Determination of trace elements in new food sources by flame atomic absorption spectrophotometry]

[Article in Chinese]

Liu LE, Ding L, Qi M, Han XL, Zhang HQ.

College of Public Health, Zhengzhou University, Zhengzhou 450052, China.

Samples were digested by HNO3 + HClO4. Flame atomic absorption spectrophotometry (FAAS) was successfully used to determine copper, zinc and iron in new resource food. Under our experimental conditions, the recovery ratio was 94.66%-108.80%; the precision was 0.71%-4.78%. This method of measuring elements is convenient, rapid and accurate. The results showed that there are profitable elements, such as copper, zinc and iron in new resourse food in Henan province. By F test and SNK test, the content sequence of metal elements was found as follows: copper, Chrysanthemum morifolium Ramat-Silktworm pupa > flowers of Pueraria lobata Ohwin = Wheat germ = Codonopsis lanceolata = roots of Pueraria lobata Ohwi > Opuntia dillenii Haw. Zinc, Opuntia dillenii Haw > Silkzowrm pupa = flowers of Pueraria lobata Ohwi = roots of Pueraria lobata Ohwi = Wheat germ = Codonopis lanceolata = Chrysanthemum morifolium Ramat. Iron, Silkworm pupa = C hrysanthemum morifolium Ramat = roots of Pueraria lobata Ohwi > flowers of Pueraria lobata Ohwi = Wheat germ = Codonopis lanceolata = Opuntia dillenii Haw.

Publication Types: PMID: 17944432 [PubMed - in process]

42. Mutational analysis of conserved outer sphere arginine residues of chalcone synthase.

Fukuma K, Neuls ED, Ryberg JM, Suh DY, Sankawa U.

Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Toyama 930-0194, Japan.

Chalcone synthase (CHS), a key enzyme in flavonoid biosynthesis, catalyses sequential decarboxylative condensations of p-coumaroyl-CoA with three malonyl-CoA molecules and cyclizes the resulting tetraketide intermediate to produce chalcone. Phenylglyoxal, an Arg selective reagent, was found to inactivate the enzyme, although no Arg is found at the active site. Conserved, non-active site Arg residues of CHS were individually mutated and the results were discussed in the context of the 3D structure of CHS. Arg199 and Arg350 were shown to provide important interactions to maintain the structural integrity and foldability of the enzyme. Arg68, Arg172 and Arg328 interact with highly conserved Gln33/Phe215, Glu380 and Asp311/Glu314, respectively, thus helping position the catalytic Cys-His-Asn triad and the (372)GFGPG loop in correct topology at the active site. In particular, a mutation of Arg172 resulted in selective impairment in the cyclization activities of CHS and stilbene synthase, a related enzyme that catalyses a different cyclization of the same tetraketide intermediate. These Arg residues and their interactions are well conserved in other enzymes of the CHS superfamily, suggesting that they may serve similar functions in other enzymes. Mutations of Arg68 and Arg328 had been found in mutant plants that showed impaired CHS activity.

Publication Types: PMID: 17938139 [PubMed - indexed for MEDLINE]

43. Major isoflavonoid contents of the 1-year-cultivated phytoestrogen-rich herb, Pueraria mirifica.

Cherdshewasart W, Sriwatcharakul S.

Department of Biology, Faculty of Science, Chulalongkorn University, Thailand. cwichai@sc.chula.ac.th

Pueraria mirifica is a tuberous plant enriched with active phytoestrogens. There is no established information about the factors influencing isoflavonoid storage in the tubers. We investigated the tuberous storage of the major isoflavonoids of 1-year-old plants. Four cultivars of P. mirifica were cultivated in the same field trial during the same period to establish a unique plant age and differentiation under the same environment and soil conditions. The tubers collected from the 1-year-old plants in the summer, rainy season and winter were submitted to an HPLC analysis with a gradient system comprising 0.1% acetic acid and acetonitrile. Five major isoflavonoids, puerarin, daidzin, genistin, daidzein and genistein, were adopted as standards. P. mirifica tubers of different cultivars collected in the same season exhibited significant differences in individual and total isoflavonoid contents, showing chemovariety. P. mirifica tubers of the same cultivar collected from different seasons also exhibited significant differences in individual and total isoflavonoid contents, showing the influence of season. In conclusion, the tuberous storage of major isoflavonoids in 1-year-cultivated plants was greatly diverse and was strongly influenced by the season and plant genetics.

Publication Types: PMID: 17928711 [PubMed - indexed for MEDLINE]

44. [Assessment of Pueraria lobata isoflavone with self-microemulsifying drug delivery systems in vitro and in vivo]

[Article in Chinese]

Cui SM, Zhao CS, He ZG.

Department of Traditional Chinese Materia Medica, Guangdong Pharmaceutical University, Guangzhou 510006, China.

OBJECTIVE: To evaluate the self-microemulsifying ability and dissolution behavior of pueraria lobata isoflavone in vitro and the pharmacokinetic behavior in rats. METHODS: The self-microemulsifying rate was evaluated by the self-microemulsifying time and the self-microemulsifying efficiency was evaluated by the particle size of resultant microemulsions. The plasma concentrations were evaluated by HPLC and dissolution and pharmacokinetic behavior of self-microemulsifying drug delivery systems were evaluated by comparison with commercial tablets. RESULTS: The system was self-microemulsified in 2 min and the particle size was less than 50 nm. The dis- solution of SMESC in distilled water was more than 90% at 10 min, while those of the commercial tablet were less than 50% at 120 min. 82% increase in the relative bioavailability was observed for the self microemulsifying drug delivery systems compared with Yufengningxin tablets. Tmax was smaller in the self-microemulsifying drug delivery systems compared with Yufengningxin tablets. CONCLUSION: The self-microemulsifying drug delivery systems can increase drug dissolution in vitro and absorption in vivo significantly.

Publication Types: PMID: 17918441 [PubMed - indexed for MEDLINE]

45. Kakkalide and irisolidone: HMG-CoA reductase inhibitors isolated from the flower of Pueraria thunbergiana.

Min SW, Kim DH.

Department of Life and Nanopharmaceutical Sciences, College of Pharmacy, Kyung Hee University, Seoul, Republic of Korea.

As part of our search for anti-arteriosclerosis agents from traditional Chinese medicines, the 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase (HCR)-inhibitory constituent, kakkalide, was isolated from the flower of Pueraria thunbergiana (PT, family Leguminosae). The antihyperlipidemic effects of kakkalide and its metabolite, irisolidone, which may be a bioactive form in vivo and potently inhibit the HCR activity, were investigated in vivo. Both the oral and interperitoneal administrations of kakkalide and irisolidone, with the exception of intraperitoneally treated kakkalide, potently lowered the serum levels of total cholesterol (TC) and triglyceride (TG) in Trition WR1339-induced hyperlipidemic mice. The oral administrations of kakkalide and irisolidone in hyperlipidemic mice induced, by the long-term feeding of a high fat diet, also potently reduced the serum levels of TC and TG and epididymal fat pad weight. These findings suggest that PT can improve hyperlipidemia, and the hypolipidemic effect may be due to HMG-CoA reductase.

PMID: 17917273 [PubMed - indexed for MEDLINE]

46. Nutrient consumption and production of isoflavones in bioreactor cultures of Pueraria lobata (Willd).

Chen G, Li L.

College of Life Science, South China Normal University, Guangdong Key Lab of Biotechnology for Plant Development, Guangzhou 510 631, China.

This paper reports the successful culture of Pueraria lobata (Willd.) suspension cells in a bioreactor. In vitro culture of this Chinese herb has potential as an alternative production method for industrial applications. Calli of P. lobata obtained from leaf explants were cultured in a 5.0 L bioreactor for two weeks. During this period, the pH of the medium declined from 5.8 to 4.5. By the end of the run about 70% of the sugars and reducing sugars and about 50% of nitrate was consumed. Almost 70% of inorganic phosphate and about 80% of the iron was exhausted. The bioreactor results indicated an isoflavone yield of 328.9 microg/ml, with an increase of about 1.77 fold. The yield of puerarin increased about 2.42 fold and reached 73.4 microg/ml in the bioreactor culture.

Publication Types: PMID: 17915773 [PubMed - indexed for MEDLINE]

47. Correlation of antioxidant activity and major isoflavonoid contents of the phytoestrogen-rich Pueraria mirifica and Pueraria lobata tubers.

Cherdshewasart W, Sutjit W.

Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Patumwan, Bangkok 10330, Thailand. cwichai@sc.chula.ac.th <cwichai@sc.chula.ac.th>

The antioxidant activity of wild Pueraria mirifica collected from 28 of the 76 provinces of Thailand and Pueraria lobata collected from China were assessed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. P. mirifica tuberous extracts showed weak antioxidant activity in comparison with alpha-tocopherol. Six plant samples exhibited stronger antioxidant activity than the mean value of the P. mirifica population. In addition, the mean value of the P. mirifica population indicated significantly lower antioxidant activity than P. lobata. The analysis of the antioxidant activity of isoflavonoids revealed that puerarin and daidzein exhibited the same level of antioxidant activity as alpha-tocopherol. The results showed convincingly that puerarin and daidzein in the plant tubers may play an important role in antioxidant activity. The correlation analysis between antioxidant activity and major isoflavonoid contents of plant tubers indicated a significant correlation only with puerarin and a significant lack of correlation with daidzin, daidzein and genistein.

Publication Types: PMID: 17890070 [PubMed - indexed for MEDLINE]

48. Protective roles of puerarin and Danshensu on acute ischemic myocardial injury in rats.

Wu L, Qiao H, Li Y, Li L.

College of Pharmaceutical Sciences, Zhejiang University, Zijingang Campus of Zhejiang University, Hangzhou 310058, China. wulimao@yahoo.com.cn

Ischemic heart diseases have been the leading cause of death in both developed and developing countries over the past decades. The aim of this study was to investigate the cardioprotective effects of the complex preparation (called Shenge), made of puerarin (isolated from Pueraria lobata Ohwi., also called Kudzu) and Danshensu (isolated from the Chinese herb, Salvia miltiorrhiza), on acute ischemic myocardial injury in rats and its underlying mechanisms. The left anterior descending (LAD) coronary artery was occluded to induce myocardial ischemia in the hearts of SD rats. Shenge was injected into the tail vein 15 min after occlusion at doses of 0, 30, 60, or 120 mg/kg body wt. ST elevation was then measured at 60, 120, and 240 min after Shenge administration. The ischemic size, serum levels of creatine kinase isoenzyme-MB (CK-MB), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA), and ST elevation were measured after the rats were sacrificed. Shenge decreased ST elevation induced by acute myocardial ischemia, reduced ischemic size, serum levels of CK-MB, LDH and MDA, and increased serum activity of SOD in a dose-dependent manner. The combined use of puerarin and Danshensu at a ratio of 1:1 showed the most effective activity. In conclusion, Shenge exerts significant cardioprotective effects against acute ischemic myocardial injury in rats, likely through its antioxidant and anti-lipid peroxidation properties, and thus may be an effective and promising medicine for both prophylaxis and treatment of ischemic heart disease.

Publication Types: PMID: 17870452 [PubMed - indexed for MEDLINE]

49. Pretreatment with phytoestrogen-rich plant decreases breast tumor incidence and exhibits lower profile of mammary ERalpha and ERbeta.

Cherdshewasart W, Panriansaen R, Picha P.

Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Patumwan, Bangkok 10330, Thailand. cwichai@sc.chula.ac.th

OBJECTIVE: Phytoestrogens have been reported to exhibit antiproliferation to human breast cancer cells in vitro. We tested the phytoestrogen-rich, Pueraria mirifica against rat breast cancer induction in vivo. METHODS: The weanling female Spargue-Dawley rats were pretreated with P. mirifica tuberous powder at a dosage of 0, 10, 100 and 1000 mg/kg BW/day for four consecutive weeks. Mammary tumor development was then induced with a single dose of 7,12-DMBA, 80 mg/kg BW, followed by a weekly examination for size and multiplicity of mammary tumors for 20 weeks and finally a necropsy. Mammary tissues were investigated for the virulence of tumor and also monoclonal antibody stained against ERalpha and ERbeta. RESULTS: Pretreatment of 1000 mg/(kgBWday) of P. mirifica tuberous powder resulted in decreasing of the virulence of rat tumor development. The mammary tumor tissues exhibited lower profile of ERalpha and ERbeta as well as ERalpha/ERbeta. CONCLUSION: P. mirifica exhibited prevention of 7,12-DMBA-induced rat mammary tumors, with a proposed mechanism of strong competitive binding of its phytoestrogens to ERalpha and/or synthesis suppressor of ERalpha.

Publication Types: PMID: 17870258 [PubMed - indexed for MEDLINE]

50. [Herbalogical textual research On "Gegen"]

[Article in Chinese]

Luo Q, Hao JD, Yang Y, Yi H.

Institute of Chinese Material Medica, China Academy of Chinese Medicinal Sciences, Beijing 100700, China.

OBJECTIVE: To provide further research with literature evidence of the variety origin, traditional effect and dosage. METHOD: Textual research on medicinal works of past dynasties. RESULT: The origin plants are Pueraria lobata, P. thomosonii and P. eduli. Its present effect is the same as ancient literature. However the fresh juice of the plant used for the treatment of bitter by mad dog and aborticide as literature recorded should be proved by further researches. The dosage is different among past dynasties. CONCLUSION: As one of the often used Chinese materia medica, Gegen has a bright prospect.

Publication Types: PMID: 17802871 [PubMed - indexed for MEDLINE]

51. Challenges in the conduct of Thai herbal scientific study: efficacy and safety of phytoestrogen, pueraria mirifica (Kwao Keur Kao), phase I, in the alleviation of climacteric symptoms in perimenopausal women.

Chandeying V, Lamlertkittikul S.

Department of Obstetrics and Gynecology, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand. verapol.c@psu.ac.th

OBJECTIVE: To evaluate the preliminary efficacy and safety of Pueraria mirifica (Kwao Keur Kao), phytoestrogen, for the alleviation of climacteric symptoms. MATERIAL AND METHOD: Perimenopausal women attending with climacteric symptoms, such as hot flushes and night sweats, were invited to join the present study, conducted at the Menopausal Clinic, Hat Yai Regional Hospital. The patients were voluntarily enrolled and randomly received the raw material of Pueraria mirifica, oral 50 and 100 mg capsule, once daily for six months, as an open-label study. RESULTS: Of the 10 enrolled patients, 8 cases were completely evaluated. The modified Greene climacteric scale (MGCS) was satisfactorily decreased in both groups. The average scale declined from 44.1 at baseline, to be 26, 17, and 11.1 at 1-, 3-, and 6- month follow-up respectively. No other laboratory abnormalities, except one case had transiently increased the creatinine level, and one case of increased blood urea nitrogen. The mean serum estradiol was slightly increased, while the mean serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were nearly stable. CONCLUSION: Pueraria mirifica is relatively safe and preliminarily alleviates the climacteric symptoms in perimenopausal. women, but the data is insufficient to draw definite conclusions regarding the estrogenic effect.

Publication Types: PMID: 17710964 [PubMed - indexed for MEDLINE]

52. [Separation and determination of puerarin, daidzin and daidzein in stems and leaves of Pueraria thomsonii by RP-HPLC]

[Article in Chinese]

Zhou HY, Wang JH, Yan FY.

College of Agronomy, Shandong Agricultural University, Taian 271018, China.

OBJECTIVE: To separation and determine the contents of puerarin, daidzin and daidzein in the stems and the leaves of Pueraria thomsonii, and to provide scientific basis for developing and using of the stems and the leaves. METHOD: A RP-HPLC method was applied with a Diamonsil C18 column (4.6 mm x 150 mm, 5 microm) by gradient elution using methanol-1% glacial acetic acid solution as the mobil phase. The flow rate was 1 mL x min(-1) and the detective wavelength was 250 nm, the column temperature was 25 degrees C. RESULT: All of the three compounds showed good linearities (r >0.9995) and the recoveries were in the range of 99.0% - 101.6%. The contents of puerarin, daidzin and daidzein in the stems are higher than those in the leaves. CONCLUSION: The method was accurate and could be used to contral the quality of the stems and leaves of P. thomsonii.

Publication Types: PMID: 17655152 [PubMed - indexed for MEDLINE]

53. Activation of large-conductance calcium-activated potassium channels by puerarin: the underlying mechanism of puerarin-mediated vasodilation.

Sun XH, Ding JP, Li H, Pan N, Gan L, Yang XL, Xu HB.

Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.

Puerarin is the main isoflavone found in Pueraria lobata (Willd) Ohwi, which has been used in therapy for various cardiovascular diseases. The present study examined the effects of puerarin on the large-conductance voltage- and Ca2+-activated potassium (BK(Ca)) channel and on rat thoracic aortas. BK(Ca) channels encoded with either alpha (BK-alpha) or alpha/beta subunits (BK-alpha+beta1) were heterologously expressed in Xenopus oocytes or human embryonic kidney 293 cells. The activities of BK(Ca) channels were measured using excised patch-clamp recordings. Puerarin activated BK-alpha+beta1 currents with a half-maximal concentration (EC50) of 0.8 nM and a Hill coefficient of 1.11 at 10 microM Ca2+ and with an EC50 of 12.6 nM and a Hill coefficient of 1.08 at 0 microM Ca2+. Puerarin (1 nM) induced a 16-mV leftward shift in the conductance-voltage curve for BK-alpha+beta1 currents at 10 microM Ca2+ and at 100 nM induced a 26-mV leftward shift at 0 microM Ca2+. Puerarin mainly increased the BK-alpha+beta1 channel open probability without changing the unitary conductance. Activation was also detected in the absence of the beta1 subunit. A deglycosylated analog of puerarin, daidzein, also activated BK(Ca) channels with weaker potency. In addition, puerarin (0.1 to 1000 microM) caused concentration-dependent relaxations of rat thoracic aortic rings contracted with 1 microM noradrenaline bitartrate (EC50 = 1.1 microM). These were significantly inhibited by 50 nM iberiotoxin, a specific blocker of BK(Ca) channels. This is the first study demonstrating that puerarin activates BK(Ca) channels, especially BK-alpha+beta1 channels. The activation of the BK(Ca) channel probably contributes to the puerarin-mediated vasodilation action.

Publication Types: PMID: 17652634 [PubMed - indexed for MEDLINE]

54. Effects of puerarin on synaptic structural modification in hippocampus of ovariectomized mice.

Xu X, Zhang Z.

College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, PR China. xuxh63@zjnu.cn

There is growing interest in whether phytoestrogens, especially isoflavones, affect cognitive function and have beneficial effects on neurodegenerative diseases. Previous work from our laboratory showed that puerarin (Pur), a major component of the Pueraria lobata isoflavones, displayed protective effects on learning and memory in ovariectomized (Ovx) female mice. In the present study, we investigated the effects of Pur on the synaptic structural modifications of the hippocampus in Ovx female mice. One week after ovariectomy or sham operation, female mice were given a 4-week treatment of Pur (50 or 100 mg/kg, I. G.) or estradiol benzoate (EB, 120 microg/kg, S. C.). The results from synaptic structural measurement and analysis did not show any differences in the hippocampal volumes, the synaptic numeric density, and the curvature of synaptic interface among Sham, Ovx, and Ovx plus EB or Pur replacement female mice. However, Pur replacement effectively normalized some structural changes induced by Ovx, reducing the width of the synaptic cleft, enlarging the thickness of postsynaptic density (PSD), and lengthening the synaptic active zone of the pre-existing synapses in the hippocampus CA1 area (P < 0.05 or P < 0.01). Furthermore, through Western blotting, we found that the immunocontent of the NMDA receptor subunit NR2 was not altered by Pur (108 % and 114 % of Ovx); however, the content of postsynaptic protein PSD-95 (145 % and 176 % of Ovx) and phosphorylation of NMDA receptor subunit NR2B (141 % and 172 % of Ovx) strongly increased in Ovx mice following treatment with Pur (50 or 100 mg/kg) for 4 weeks. These results suggest that Pur possesses phytoestrogen activity with 1,000 -fold lower activity than EB, and the beneficial effects of Pur on improving memory behavior of Ovx female mice are associated with the changes of synaptic structural modifications in the hippocampus.

Publication Types: PMID: 17628835 [PubMed - indexed for MEDLINE]

55. Assessment of fertility and reproductive toxicity in adult female mice after long-term exposure to Pueraria mirifica herb.

Jaroenporn S, Malaivijitnond S, Wattanasirmkit K, Watanabe G, Taya K, Cherdshewasart W.

Interdepartment of Physiology, Faculty of Graduate School, Chulalongkorn University, Bangkok, Thailand.

The present study investigated the effects of long-term administration of Pueraria mirifica (PM) at non-toxic doses on the ovarian function and fertility of adult female mice based on evaluation of hematological and biochemical parameters. Female mice were divided into 4 groups (36 mice/group). Groups 1-3 were orally treated with a dose of 0 (PM-0), 10 (PM-10) or 100 mg/kg BW/day PM (PM-100), and group 4 was subcutaneously injected with 200 mug/kg BW/day of synthetic estrogen diethylstilbestrol (DES). The treatment schedule was separated into treatment and post-treatment periods. The duration of each period was 8 weeks. The PM-10 mice exhibited regular estrous cycles, while the PM-100 and DES treatments induced prolonged estrous cycles. Although no changes were observed in the uterus and ovary weights of the mice after the PM-100 and DES treatments, hyperplasia of the uterine endothelium and a decrease in the number of growing ovarian follicles were detected. The changes in the ovarian histologies of the PM-100 and DES mice were related to reductions in the levels of LH and FSH, which subsequently caused a decrease in mating efficiency. Once the PM mice were able to copulate, they were capable of successfully becoming pregnant and mothering offspring. No abnormalities were observed in the external morphologies and reproductive organ weights of the 50-day-old offspring. In conclusion, our results suggest that long-term exposure to 100 mg/kg BW of PM has adverse effects on the mating efficiency and reproduction of adult female mice and that administration of 10 mg/kg BW of PM does not induce any changes in the hypothalamic-pituitary-ovarian-uterine axis.

Publication Types: PMID: 17585183 [PubMed - indexed for MEDLINE]

56. Effect of serum metabolites of Pueraria lobata in rats on peripheral nerve regeneration: in vitro and in vivo studies.

Chen HT, Yao CH, Chao PD, Hou YC, Chiang HM, Hsieh CC, Ke CJ, Chen YS.

Laboratory of Biomaterials, Graduate Institute of Chinese Medical Science, China Medical University, Taichung, Taiwan.

This study provides in vitro and in vivo evaluation of rat serum metabolites of the Pueraria lobata (SMP) on peripheral nerve regeneration. In the in vitro study, we found that the SMP caused a marked enhancement of the nerve growth factor (NGF)-mediated neurite outgrowth and the expression of synapsin I from PC12 cells. In the in vivo study, silicone rubber chambers filled with the SMP were used to bridge a 10-mm sciatic nerve defect in rats. At the conclusion of 8 weeks, animals from the groups treated with the SMP had a relatively more mature structure with larger mean values of myelinated axon number, endoneurial area, and total nerve area when compared with those in the controls receiving the saline only. These results suggest that the serum metabolites of Pueraria lobata can be a potential nerve growth-promoting factor.

Publication Types: PMID: 17514670 [PubMed - indexed for MEDLINE]

57. Seasonal variations in the isoflavonoids of radix Puerariae.

Sibao C, Dajian Y, Shilin C, Hongx X, Chan AS.

State Key Laboratory of Chinese Medicine and Molecular Pharmacology, Shenzhen, 518057, People's Republic of China.

The variations in isoflavonoid content in Radix Puerariae, the root of Pueraria lobata (Wild) Ohwi, have been examined in order to determine the optimum time to harvest the herb. Total isoflavonoid contents have been analysed by a UV spectrophotometric method whilst HPLC with photodiode array detection has been used to monitor the contents of seven major individual isoflavonoids, namely, 3'-hydroxypuerarin, puerarin, 3'-methoxypuerarin, daidzin, genistin, formononetin-7-glucoside and daidzein. Ninety-six samples of Radix Puerariae of different ages and harvested in different months were collected and analysed. The results clearly showed that 3-year-old roots harvested in January have the highest yields of isoflavonoid compounds. Moreover, the results obtained by both methods have a strong correlation. This suggested that the simple and fast UV spectrophotometric method could be used for monitoring the phytochemical quality of Radix Puerariae.

Publication Types: PMID: 17500368 [PubMed - indexed for MEDLINE]

58. Cytoprotective effects of KIOM-79 on streptozotocin induced cell damage by inhibiting ERK and AP-1.

Kang KA, Lee KH, Kim SY, Kim HS, Kim JS, Hyun JW.

Department of Biochemistry, College of Medicine and Applied Radiological Science Research Institute, Cheju National University, Jeju-Si, Korea.

The present study investigated the potential cytoprotective properties of a combination of plant extracts (KIOM-79) obtained from Magnolia officinalis, Pueraria lobata, Glycyrrhiza uralensis, and Euphorbia pekinensis, against the oxidative stresses induced by streptozotocin (STZ) in a rat pancreatic beta-cells (RINm5F). KIOM-79 was found to scavenge intracellular reactive oxygen species (ROS), thereby preventing DNA damage and lipid peroxidation. The KIOM-79 inhibited apoptosis of the beta-cells exposed to STZ via radical scavenging activity and activation of antioxidant enzymes. KIOM-79 inhibited activation of extracellular regulated kinase (ERK) induced by STZ and inhibited DNA binding activity of an activator protein-1 (AP-1), a downstream transcription factor of ERK. Taken together, these findings suggest that KIOM-79 protects against STZ induced cell death in RINm5F cells by inhibiting ROS generation and the ERK pathway.

Publication Types: PMID: 17473425 [PubMed - indexed for MEDLINE]

59. Stimulatory effect of puerarin on bone formation through activation of PI3K/Akt pathway in rat calvaria osteoblasts.

Zhang Y, Zeng X, Zhang L, Zheng X.

Department of Biomedical Engineering, Key Laboratory of Biomedical Engineering of Ministry of Education, Zhejiang University, Hangzhou, P. R. China.

Puerarin, a natural isoflavonoid found in Chinese Pueraria lobata (Wild.) Ohwi, has received increasing attention because of its possible role in the prevention of osteoporosis. However, the relationship between puerarin and bone formation remains unknown. In the present study, rat osteoblasts isolated from newborn Wistar rats were used to investigate the effect of puerarin on osteoblasts, and its possible molecular mechanism. Data showed that puerarin caused a significant increase in cell viability, alkaline phosphatase (ALP) activity and mineral nodules formation in osteoblasts, suggesting that puerarin had a stimulatory effect on osteoblastic bone formation. This functional improvement by puerarin was accompanied by activation and nuclear translocation of Akt. Furthermore, puerarin-stimulated osteoblastic growth, Akt activation and redistribution were significantly blocked by the specific PI3K inhibitor, LY294002. These results strongly suggested that puerarin stimulated osteoblastic proliferation and Akt activation in a PI3K-dependent manner. In summary, puerarin derived from Chinese Pueraria lobata (Wild.) Ohwi can promote bone formation in cultured rat osteoblasts, which might be mediated by activation of the PI3K/Akt pathway. DMEM:Dulbecco's modification of Eagel's medium PBS:phosphate buffered saline DMSO:dimethyl sulfoxide EDTA:ethylene diamine tetraacetic acid SDS:sodium dodecyl sulfate SDS-PAGE:sodium dodecylsulfate polyacrylamide gel electrophoresis FITC:fluorescein isothiocyanate HRP:horseradish peroxidase PI3K:phosphatidylinositol 3-kinase.

Publication Types: PMID: 17443435 [PubMed - indexed for MEDLINE]

60. Effect of Pueraria mirifica on vaginal health.

Manonai J, Chittacharoen A, Theppisai U, Theppisai H.

Department of Obstetrics and Gynaecology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand. rajmo@mahidol.ac.th

OBJECTIVE: To evaluate the effect of Pueraria mirifica on vaginal symptoms, vaginal health index, vaginal pH, and vaginal cytology in healthy postmenopausal women. DESIGN: A randomized, double-blind, placebo-controlled study. Healthy postmenopausal women, age 45 to 60 years old, were enrolled voluntarily and randomly received 20, 30, or 50 mg of Pueraria mirifica in capsules or placebo in identical capsules once daily for 24 weeks. RESULTS: After 24 weeks of treatment, 71 women were evaluated. Fifty-one of 71 randomly received one of the three doses of Pueraria mirifica, and the remaining 20 received placebo. The mean vaginal dryness symptom in the Pueraria mirifica group decreased after 12 weeks of treatment. Pueraria mirifica increased vaginal maturation index (parabasal:intermediate:superficial cells) from 46:43:11 to 11:65:24 after 24 weeks of treatment. There was no significant difference of adverse effects between the Pueraria mirifica and placebo groups in this study. CONCLUSIONS: Pueraria mirifica was proven to exhibit estrogenicity on vaginal tissue, to alleviate vaginal dryness symptoms and dyspareunia, to improve signs of vaginal atrophy, and to restore the atrophic vaginal epithelium in healthy postmenopausal women.

Publication Types: PMID: 17415017 [PubMed - indexed for MEDLINE]

61. Suitability of Pueraria phaseoloides, Chromolaena odorata and Tithonia diversifolia as in-situ mulch for nematode management in musa cropping systems.

Schösser B, Hauser S, Sikora RA.

Nematology in Soil Ecosystems, INRES--Institute for Crop Science and Resource Conservation, University of Bonn, Nussallee 9, DE-53115 Bonn, Germany. birthe.schoesser@web.de

Mulching with plant organic matter has been shown to reduce nematode population densities in various cropping systems. The level of nematode control is increased when such mulches are incorporated into the soil as organic amendments. Chromolaena odorata, Tithonia diversifolia and Pueraria phaseoloides are common cover crops in West and Central Africa that produce large quantities of nutrient rich biomass. The aim of this study was to determine, if in-situ mulching of C. odorata, T. diversifolia and P. phaseoloides is suitable for nematode control in Musa production. In a pot trial, the susceptibility of these plants to spiral nematodes was investigated. The effects of different quantities of surface mulch on nematode population densities in the soil and in banana roots also were determined. All mulch types and all quantities led to a reduction in nematode population densities in the soil. The strongest nematode reductions were observed in the Pueraria treatments. In treatments containing banana plants mulching improved plant growth compared to the clean-fallowed soil and induced lower root infestation rates. However, nematode soil populations were higher in mulched than in non-mulched banana treatments. Plant parasitic nematodes also were isolated from roots of all three cover crop species and all three plants caused an increase in nematode numbers in the soil. Therefore, the tested cover crops proved unsuitable for nematode control in a system with the highly susceptible bananas. Further examinations are needed to determine whether or not the positive effects of surface mulching on plantain plant growth and root infestation rates also have positive effects on yield in an in-situ mulching system in the presence of nematodes.

PMID: 17390809 [PubMed - indexed for MEDLINE]

62. Effect of puerarin on bone formation.

Wong R, Rabie B.

University of Hong Kong, Department of Orthodontics, Prince Philip Dental Hospital, Hong Kong. fyoung@hhkucc.hku.hk

OBJECTIVE: Puerarin is one of the major phytoestrogens isolated from Pueraria lobata, a Chinese medicine known as Gegen. Our laboratory compared the amount of new bone produced by puerarin in collagen matrix (carrier) to that produced by the collagen matrix alone. METHOD: Eighteen bone defects, 5mm by 10mm were created in the parietal bone of nine New Zealand White rabbits. In the experimental group, six defects were grafted with puerarin solution mixed with collagen matrix. In the control groups, six defects were grafted with collagen matrix alone (active control) and six were left empty (passive control). Animals were killed on day 14 and the defects were dissected and prepared for histological assessment. Serial sections were cut across each defect. No new bone was formed in the passive control group. Quantitative analysis of new bone formation was made on 100 sections (10 sections in each defect, in five defects randomly selected in each of the experimental group and active control group) using image analysis. RESULTS: A total of 554% more new bone was present in defects grafted with puerarin in collagen matrix than those grafted with the collagen matrix alone. CONCLUSION: Puerarin in collagen matrix has the effect of increasing new bone formation locally and can be used for bone grafting or for bone induction often required in surgery.

Publication Types: PMID: 17379543 [PubMed - indexed for MEDLINE]

63. Identification of isoflavone glycosides in Pueraria lobata cultures by tandem mass spectrometry.

Prasain JK, Reppert A, Jones K, Moore DR 2nd, Barnes S, Lila MA.

Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, AL 35294, USA. jprasain@uab.edu

Isoflavones in the methanolic extracts of kudzu (Pueraria lobata) callus, suspension and root cultures were compared in order to develop an experimental system in which puerarin (daidzein 8-C-glucoside) and other isoflavones could be synthesised in vitro. Quantitative variation of puerarin and other known isoflavones was estimated in kudzu culture extracts using HPLC-UV. The highest and lowest amounts of puerarin (14.56 and 0.33 mg/g) were present in in vitro root cultures and leaf tissue-derived callus cultures, respectively. A total of 48 isoflavone metabolites were detected in extracts of kudzu root cultures by HPLC-MS/MS, and the structures of 33 of them were tentatively assigned. Amongst these, 12 isoflavone C-glycosides were identified. Hydroxyderivatives of puerarin in several isomeric forms were detected, some of which have not been previously reported in kudzu root. The molecular weights, interpretation of characteristic fragment ions obtained from HPLC-MS/MS and comparison with reported data allowed the putative identification of the isoflavone metabolites.

Publication Types: PMID: 17260698 [PubMed - indexed for MEDLINE]

64. Identification of isoflavones in the roots of Pueraria lobata.

Rong H, Stevens JF, Deinzer ML, Cooman LD, Keukeleire DD.


University of Gent, Faculty of Pharmaceutical Sciences, Gent, Belgium.

The isoflavones of the roots of Pueraria lobata (Willd.) Ohwi (Puerariae Radix) were investigated by high-performance liquid chromatography (HPLC) coupled to photodiode array (PDA) and to mass spectroscopy (MS) using atmospheric pressure chemical ionization (APCI) or electrospray ionization (ESI) in combination with collision-activated decomposition (CAD) (HPLC-APCI-CAD-MS or ESI-CAD-MS) for identification of glycosides and HPLC-APCI-CAD-MS for identification of aglycones. The major glycosides are derived from daidzein ( 9) and most are 8- C-glycosides. 3'-Hydroxypuerarin-4'- O-deoxyhexoside ( 2B) and 3'-methoxy-6''- O- D-xylosylpuerarin ( 6) were identified as new constituents. MS data were obtained for puerarin-4'- O- D-glucoside ( 1), 3'-hydroxypuerarin ( 2A), puerarin ( 3), 3'-methoxypuerarin ( 4), 6''- O- D-xylosylpuerarin ( 5), daidzin ( 7) and 3'-methoxydaidzin ( 8), which were previously characterized by NMR analysis. Isoflavones identified in Puerariae Radix comprise 3'-methoxydaidzein ( 10), genistein ( 12), daidzein-7- O-methyl ether ( 13A), 3'-methoxydaidzein-7- O-methyl ether or 3'-methoxyformononetin ( 13B) and biochanin A ( 15), while previous characterization of daidzein ( 9) and formononetin ( 14) was substantiated by MS data. The structure of compound 11 could not be established by MS techniques. The estrogenic activity was mainly located in the aglycone fraction.

PMID: 17253303 [PubMed]

63. Evaluation of the estrogenic activity of the wild Pueraria mirifica by vaginal cornification assay.

Cherdshewasart W, Kitsamai Y, Malaivijitnond S.

Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand. cwichai@sc.chula.ac.th

The aim of this study was to evaluate the estrogenic activity of tuberous samples of phytoestrogen-rich Pueraria mirifica collected from 25 of 76 provinces in Thailand by vaginal cornification assay. Tuberous powders were prepared and administered to ovariectomized rats for 14 consecutive days at dosages of 10, 100 and 1,000 mg/kg BW respectively, and were compared with a daily treatment with 2 mg/kg BW 17beta-estradiol (E(2)). Rats treated with 10 mg/kg BW Pueraria mirifica showed no vaginal cornification. Treatment with 100 mg/kg BW Pueraria mirifica from 13 out of 25 plant samples resulted in development of vaginal cornification. The cell count percentages of the vaginal smeared cells for the treatment with the 2 plant samples that exhibited the fastest vaginal cornification revealed large variation in their estrogenic activities. Treatment with 1,000 mg/kg BW Pueraria mirifica from all plant samples produced vaginal cornification with the mean value for the period (day) of first appearance of cornified cells being 4.08 days compared to 2 days with 2 mg/kg BW E(2). The overall appearance period (day) of cornified cells during the treatment and post-treatment period with 1,000 mg/kg BW per day Pueraria mirifica was shorter than treatment with 2 mg/kg BW E(2). The results demonstrate that the plant population shows differential estrogenic activity as evaluated by vaginal cornification assay.

Publication Types: PMID: 17229996 [PubMed - indexed for MEDLINE]

66. Assessment of urinary gonadotropin and steroid hormone profiles of female cynomolgus monkeys after treatment with Pueraria mirifica.

Trisomboon H, Malaivijitnond S, Cherdshewasart W, Watanabe G, Taya K.

Primate Research Unit, Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.

This study investigated the changes in the urinary hormone levels of female monkeys (Macaca fascicularis) after single-dose and long-term treatments with Pueraria mirifica (PM). The monkeys were separated into 3 groups (n=3) and orally treated with 10, 100, or 1,000 mg of PM in each group. Two series of experiments were performed. In the first series of experiments, the monkeys were orally treated with a single dose of PM. The experimental schedule was divided into a one menstrual cycle pretreatment period and a two menstrual cycle post-treatment period. In the second series of experiments, the monkeys were orally treated daily with PM for 90 days. The experiment schedule was divided into a one menstrual cycle pretreatment period, a three menstrual cycle treatment period, and a two menstrual cycle post-treatment period. Urinary samples were collected daily and assayed for the FSH, LH, estradiol, and progesterone levels. The results showed that there were no changes in the FSH, LH, estradiol, and progesterone levels after treatment with a single dose of 10, 100, or 1,000 mg of PM or after daily treatment with 10 mg of PM for 90 days compared with the levels observed during the pretreatment period. Daily treatment with 100 mg and 1,000 mg of PM for 90 days only produced a clear reduction in the urinary FSH levels. This suggests that changes of urinary FSH levels can be considered an indicator for study of estrogenic effects on hormonal levels in female monkeys.

Publication Types: PMID: 17202751 [PubMed - indexed for MEDLINE]

67. Molecular authentication of Radix Puerariae Lobatae and Radix Puerariae Thomsonii by ITS and 5S rRNA spacer sequencing.

Sun Y, Shaw PC, Fung KP.

Department of Biochemistry and Institute of Chinese Medicine, The Chinese University of Hong Kong, NT.

In the present study, we examined nuclear DNA sequences in an attempt to reveal the relationships between Pueraria lobata (Willd). Ohwi, P. thomsonii Benth., and P. montana (Lour.) Merr. We found that internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA are highly divergent in P. lobata and P. thomsonii, and four types of ITS with different length are found in the two species. On the other hand, DNA sequences of 5S rRNA gene spacer are highly conserved across multiple copies in P. lobata and P. thomsonii, they could be used to identify P. lobata, P. thomsonii, and P. montana of this complex, and may serve as a useful tool in medical authentication of Radix Puerariae Lobatae and Radix Puerariae Thomsonii.

Publication Types: PMID: 17202681 [PubMed - indexed for MEDLINE]

68. Anti-inflammatory mechanisms of isoflavone metabolites in lipopolysaccharide-stimulated microglial cells.

Park JS, Woo MS, Kim DH, Hyun JW, Kim WK, Lee JC, Kim HS.

Department of Neuroscience, College of Medicine, Ewha Womans University, Mok-6-dong 911-1, Yangchun-Ku, Seoul 158-710, South Korea.

The microglial activation plays an important role in neurodegenerative diseases by producing several proinflammatory cytokines and nitric oxide (NO). We found that three types of isoflavones and their metabolites that are transformed by the human intestinal microflora suppress lipopolysaccharide (LPS)-induced release of NO and tumor necrosis factor (TNF)-alpha in primary cultured microglia and BV2 microglial cell lines. The inhibitory effect of the isoflavone metabolites (aglycon form) was more potent than that of isoflavones (glycoside form). The RNase protection assay showed that the isoflavone metabolites regulated inducible nitric oxide synthase (iNOS) and the cytokines at either the transcriptional or post-transcriptional level. A further molecular mechanism study was performed for irisolidone, a metabolite of kakkalide, which had the most potent anti-inflammatory effect among the six isoflavones tested. Irisolidone significantly inhibited the DNA binding and transcriptional activity of nuclear factor (NF)-kappaB and activator protein-1. Moreover, it repressed the LPS-induced extracellular signal-regulated kinase (ERK) phosphorylation without affecting the activity of c-Jun N-terminal kinase or p38 mitogen-activated protein kinase. The level of NF-kappaB inhibition by irisolidone correlated with the level of iNOS, TNF-alpha, and interleukin (IL)-1beta suppression in LPS-stimulated microglia, whereas the level of ERK inhibition correlated with the level of TNF-alpha and IL-1beta repression. Overall, the repression of proinflammatory cytokines and iNOS gene expression in activated microglia by isoflavones such as irisolidone might have therapeutic potential for various neurodegenerative diseases including ischemic cerebral disease.

Publication Types: PMID: 17194798 [PubMed - indexed for MEDLINE]

69. Effects of Pueraria mirifica, an herb containing phytoestrogens, on reproductive organs and fertility of adult male mice.

Jaroenporn S, Malaivijitnond S, Wattanasirmkit K, Trisomboon H, Watanabe G, Taya K, Cherdshewasart W.

Interdepartment of Physiology, Faculty of Graduate School, Chulalongkorn University, Bangkok 10330, Thailand.

The effects of Pueraria mirifica (PM) on reproductive organs and fertility of adult male mice were investigated. Male mice were divided into four groups (10 mice/group). Groups 1-3 were orally treated with PM at doses of 0 (PM-0), 10 (PM-10), and 100 (PM-100) mg/kg BW/d in 0.2 mL distilled water, and group 4 was subcutaneously injected with 200 microg/kg BW/d of synthetic estrogen diesthylstilbestol (DES). The treatment schedule was separated into two periods: treatment and posttreatment (8 wk for each period). The PM-10 and PM-100 treatments had no effect on testicular weight, sperm number, and serum LH, FSH, and testosterone levels. Only the PM-100 treatment reduced weights of epididymes and seminal vesicle and the sperm motility and viability. Histopathological examination demonstrated that testis, epididymis, and seminal vesicle were normal in all doses of PM treatment. PM-treated males showed no alterations in mating efficiency and on causing pregnancy of their female partners. DES injection impaired all those parameters. Offspring fathered by the PM- and DES-treated males exhibited neither malformations nor change of body weight gains, and the reproductive organ weights of 50-d old pups were in the normal range. The present data clearly demonstrate that a long-term treatment of PM at doses 10 and 100 mg/kg BW/d, via oral route, does not alter a male fertility and a hypothalamus- pituitary-testis axis. Although PM-100 can cause some moderate impairment, no persistent effects were observed. Most of PM-treated mice increased the mating efficiency after stop treatment.

Publication Types: PMID: 17185797 [PubMed - indexed for MEDLINE]

70. Isoflavonoids production in callus culture of Pueraria tuberosa, the Indian kudzu.

Vaishnav K, Goyal S, Ramawat KG.


Laboratory of Biomolecular Technology, Department of Botany, M L Sukhadia University, Udaipur 313 001, India.

Isoflavonoid contents of different plant parts and callus tissues of the Indian Kudzu, Pueraria tuberosa (Roxb.ex.Willd.) DC are presented. The initial cultures were slow growing, associated with browning of the tissues. The production of four isoflavonoids (puerarin, genistin, genistein and daidzein) in the callus cultures of P. tuberosa was studied by manipulating the plant growth regulators and sucrose concentration in the medium. Organogenesis was not recorded in callus on any of these treatments. Tuber and stem accumulated puerarin, a glycoside of daidzein, at high amounts, 0.65% and 0.054% respectively. However, the daidzein content of the callus tissues grown on Murashige and Skoog medium containing BA (20.9 microM) and sucrose (60 gl(-1)) was significantly higher (0.056%) than in vivo plant material (0.02%) and other comparable culture systems like Genista and Pueraria lobata.

Publication Types: PMID: 17176676 [PubMed - indexed for MEDLINE]

71. [Effect of several physiochemical factors on cell growth and isoflavone accumulation of Pueraria lobata cell suspension culture]

[Article in Chinese]

Fang CB, Li HQ, Wan XC, Jiang CJ.

Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China.

OBJECTIVE: To illustrate the effects of several physiochemical factors on cell growth and isoflavone accumulation of Pueraria lobata cell suspension cultures. METHOD: High performance liquid chromatography and plant tissue culture were applied. RESULT AND CONCLUSION: Cell growth and isoflavone accumulation were significantly stimulated in P. lobata cell suspension cultures by the increase of the sucrose concentration. Maintaining the pH value at the range over 5. 4 to 5. 8 was most suitable for isoflavone accumulation in P. lobata cell suspension cultures. Cell dried weight and isoflavone accumulation decreased sharply with the increase of the treated concentration of active carbon, while XAD-4 significantly stimulated cell growth and isoflavone accumulation.

Publication Types: PMID: 17165578 [PubMed - indexed for MEDLINE]

72. A Lupinoside prevented fatty acid induced inhibition of insulin sensitivity in 3T3 L1 adipocytes.

Dey D, Pal BC, Biswas T, Roy SS, Bandyopadhyay A, Mandal SK, Giri BB, Bhattacharya S.

Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Kolkata 700032, India.

The decrease in insulin sensitivity to target tissues or insulin resistance leads to type 2 diabetes mellitus, an insidious disease threatening global health. Numerous evidences made free fatty acids (FFAs) responsible for insulin resistance and type 2 diabetes. We demonstrate here that the damage of insulin acitivity by a free fatty acid, palmitate could be prevented by a lupinoside. An incubation of 3T3 L1 adipocytes with a FFA i.e. palmitate inhibited insulin stimulated uptake of (3)H-2 deoxyglucose (2 DOG) significantly. Addition of a lupinoside purified from Pueraria tuberosa, lupinoside PA(4) (LPA(4)) strongly prevented this inhibition. We then examined insulin signaling pathway where palmitate significantly inhibited insulin stimulated phosphorylation of Insulin receptor tyrosine kinase, IRS 1and PI3 kinase, PDK1 and Akt/PKB. LPA(4) rescued this inhibition of signaling molecule by palmitate. Insulin mediated translocation of Glut4, the glucose transporter in insulin target cells, was effectively blocked by palmitate while, LPA(4) waived this block. Administration of LPA(4) to nutritionally induced diabetic rats significantly reduced the increase in plasma glucose. All these indicate LPA(4) to be a potentially therapeutic agent for insulin resistance and type 2 diabetes.

PMID: 17149545 [PubMed - indexed for MEDLINE]

73. Intestinal bacteria activate estrogenic effect of main constituents puerarin and daidzin of Pueraria thunbergiana.

Park EK, Shin J, Bae EA, Lee YC, Kim DH.

College of Pharmacy, Kyung Hee University, Seoul, Korea.

To understand the relationship between the metabolites and estrogenic activity of the main isoflavones puerarin and daidzin of the rhizome of Pueraria thunbergiana (PT, family Leguminosae), PT and its isoflavones were transformed by human intestinal bacteria and their estrogenic effects were investigated. All human fecal specimens hydrolyzed puerarin and daidzin to daidzein, but their hydrolyzing activities varied depending on the individuals. All intestinal bacteria isolated from human also hydrolyzed daidzin to daidzein, but a few bacteria transformed puerarin to daidzein. When the estrogenic effect of PT, puerarin and daidzin was compared with those of their metabolites, the metabolites more potently increased proliferation of MCF-7 cells than PT, puerarin and daidzin. The metabolite daidzein also potently increased estrogen-response c-fos mRNA and PR protein expressions. These findings suggest that intestinal bacteria, which can hydrolyze puerarin and/or daidzin, may activate a potent estrogenic activity of PT.

Publication Types: PMID: 17142977 [PubMed - indexed for MEDLINE]

74. Constituents of the roots of Pueraria lobata inhibit formation of advanced glycation end products (AGEs).

Kim JM, Lee YM, Lee GY, Jang DS, Bae KH, Kim JS.

Department of Herbal Pharmaceutical Development, Korea Institute of Oriental Medicine, Daejeon 305-811, Korea.

Two isoflavone C-glucosides, puerarin (1) and PG-3 (2), a but-2-enolide, (+/-)-puerol B (3), two isoflavone O-glucosides, daidzin (4) and genistin (5), and three pterocarpans, (-)-medicarpin (6), (-)-glycinol (7) and (-)-tuberosin (8), were isolated from a MeOH extract of the roots of Pueraria lobata, using an in vitro bioassay based on the inhibition of the formation of advanced glycation end products (AGEs) to monitor chromatographic fractionation. The structures of 1-8 were determined by spectroscopic data interpretation, particularly by 1D- and 2D-NMR studies, and by comparison of these data with values in the literature. All of the isolates (1-8) were evaluated for their inhibitory activity on AGEs formation in vitro. Of these, puerarin (1), PG-3 (2), and (+/-)-puerol B (3) exhibited more potent inhibitory activity than the positive control aminoguanidine.

Publication Types: PMID: 17121174 [PubMed - indexed for MEDLINE]

75. Pueraria mirifica, a phytoestrogen-rich herb, prevents bone loss in orchidectomized rats.

Urasopon N, Hamada Y, Asaoka K, Cherdshewasart W, Malaivijitnond S.

Biological Science Ph.D. Program, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.

OBJECTIVE: Estrogens and estrogen-like substances have been reported to play an important role in male bone homeostasis and to prevent bone loss. Pueraria mirifica (Leguminosae), a Thai herbal plant, containing a high amount of phytoestrogens was a choice of interest for this study. We examined the effects of crude P. mirifica on bone loss and influences on reproductive organs in male rats. METHODS: Using fully mature and orchidectomized (ORX) rats, the effects of 0, 10, 100 and 1000 mg/kgB.W./day of P. mirifica and 0.1mg/kg B.W./day of 17 alpha-ethinylestradiol (a positive control) were evaluated on bone mineral density (BMD) and bone mineral content (BMC) measured with a peripheral Quantitative Computerized Tomography (pQCT) densitometry. RESULTS: Bone loss in trabecular and cortical bones of the various sites of axial bone (fourth lumbar vertebral body) and long bones (tibia and femur) after ORX was dose-dependently prevented by P. mirifica. The effects were specific on bone types and sites. The weights of the accessory sex organs, seminal vesicle and ventral prostrate gland, which significantly decreased after 3-month of ORX, were not altered by P. mirifica. CONCLUSION: The results suggest that P. mirifica treatment may be useful to prevent an osteoporosis in elderly hypogonadism subjects without influences on reproductive organs.

Publication Types: PMID: 17101247 [PubMed - indexed for MEDLINE]

76. [Preparation of puerarin solid self-microemulsion]

[Article in Chinese]

Yu AH, Zhai GX, Cui J, Liu H.

College of Pharmacy, Shandong University, Jinan 250012, China. yuaihua@sdu.edu.cn

OBJECTIVE: To prepare puerarin solid self-microemulsifying drug delivery system (SMEDDS). METHODS: Pseudo-ternary phase diagram was used to select the formula of microemulsion. The self-microemulsification efficiency was assessed, such as viscosity, conductance, refraction, zeta potential, size ditribution, dissolution were investigated. Puerarin were determined by UV to calculate the cumulative release at different time. RESULTS: The optimum formulation of puerarin solid self-microemulsifying drug delivery system the consisted of puerarin-Tween 800 glycerinum-ethyl oleate-water-Gum Acacia-manicol at weight ratio of 1.7: 16. 6: 16. 6: 16. 6 : 1.3: 4.0: 39.9: 19.9; Average particle diameter was 30 nm; The dissolution was 94.29% at 10 min. CONCLUSION: Stability of puerarin solid self-microemulsion drug delvery system is good, potent in improving the dissolution of puerarin.

Publication Types: PMID: 17076244 [PubMed - indexed for MEDLINE]

77. [Studies on the effect of extracts of several Chinese herbal medicines and other medicines on alcohol dehydrogenase activity]

[Article in Chinese]

Li WZ, Lu J, Sun XY, Wang SL, Ni YM, Zhu WS.

College of Life Science and Technology of Tongji University, Shanghai 200092, China. lwz@mail.tongji.edu.cn

OBJECTIVE: To study the effects of water and alcohol extracts of several Chinese herbal medicines and other medicines on alcohol dehydrogenase activity in order to provide enzymology basis on new medicine. METHODS: Water or alcohol extracts of Chinese herbal medicine and other medicine were tested on the effects of alcohol dehydrogenase activity by Valle and Hoch method. RESULTS: Among them, 8 were found to have the effect of activation on alcohol dehydrogenase. They were water extracts of Amomum kravanh and Pueraria flowers, the alcohol extracts of Pueraria flowers, compound hepatcare Chinese medicine and compound Pueraria medicine, L-cysteine, notoginseng saponin. Others had inhibiting action. CONCLUSION: To decrease alcohol concentration in the body through activating the activity of ADH may be one of the mechanisms for some traditional Chinese herbal medicine in neutralizing the effect of alcohol drink.

Publication Types: PMID: 17076243 [PubMed - indexed for MEDLINE]

78. [Improvement effects of puerarin on glycated brain damages in rats induced by D-galactose]

[Article in Chinese]

Lv JH, Zhang SP, Shen FH, Pan JQ, Tan HR.

School of Pharmacy, Jinan University, Guangzhou 510632, China. yaolilv@163.com

OBJECTIVE: To observe the improvement effects of puerarin on glycated brain damages in rat model induced by D-galactose. METHOD: The model rats of protein glycation were induced by intraperitoneal administration of D-galactose (150 mg x kg(-1) x d(-1)) for 8 weeks, and all rats were treated with puerarin (high dose 300 mg x kg(-1), middle dose 150 mg x kg(-1), low dose 75 mg x kg(-1)) for 6 weeks. The activity of aldose reductase in red blood cells, the amount of glycated products (fructosamine in serum, glycohaemoglobin, advanced glycation end-products) and AGEs in brain tissue, calcium ion in brain cells were measured. Moreover, mitochondria in brain hippocampus cells were observed under electronic microscope. RESULT: High dose and middle dose of puerarin can decrease the activity of aldose reductase in red blood cells (P < 0.01), and inhibit the formation of glycation products significantly in model rats induced by D-galactose (P < 0.01). Also, puerarin can decrease the content of AGEs in brain and the level of calcium ions in brain cells (P < 0.05, P < 0.01), and decrease lesions degree in mitochondria in brain hippocampus cells. CONCLUSION: Puerarin can produce the protective effects on glycated brain damages through inhibiting the glycation reaction in rats induced by D-galactose.

Publication Types: PMID: 17048591 [PubMed - indexed for MEDLINE]

79. [Primary study on acid tolerance mechanism of a wild aciduric Rhizobium strain isolated from Pueraria lobata]

[Article in Chinese]

Gu JP, Zhang L, Wei SQ, Zhang Q, Fang DH.

College of Resource and Environment, Southwest University, Chongqing 400716, China. gwping80@163.com

An aciduric Rhizobium strain, named as PR389, was isolated from the nodule of wild Pueraria lobata which grew in yellow soil (pH 4.6) on the Jin-yun Mountain in Bei-bei, Chong-qing city. The isolated strain, which could grow under pH 4.6 distinct from the optimal pH 6.5-7.5 for rhizobium, showed some typical features of aciduric rhizobium. This was also confirmed by the proton flux assay. Compared to the acid-sensitive Rhizobium strain PR21, the cell membrane of PR389 could hold back excessive H entering cell. This feature can protect PR389 from harm of acid. In the test of acid tolerance, the aciduric ability of strain PR389 under low acidic (pH 3.8) was restrained by antibiotic chloramphenicol. It was speculated that special proteins in the cells of PR389 could be induced and synthesized in acidic environment.

Publication Types: PMID: 17037073 [PubMed - in process]

80. Puerarin, an isoflavonoid derived from Radix puerariae, potentiates endothelium-independent relaxation via the cyclic AMP pathway in porcine coronary artery.

Yeung DK, Leung SW, Xu YC, Vanhoutte PM, Man RY.

Department of Pharmacology, 2/F Faculty of Medicine Building, University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong, People's Republic of China.

Puerarin, an isoflavonoid derived from the Chinese medicinal herb Radix puerariae, has been suggested to be useful in the management of various cardiovascular disorders. The present study examined the effect of acute exposure (30 min) to puerarin on vascular relaxation. Rings from porcine coronary artery of either sex were used. The highest concentration of puerarin (100 microM) produced a small but statistically significant relaxation of U46619-contracted rings. Vascular relaxations were also studied in the presence of lower concentrations of puerarin (0.1, 1 and 10 microM) which had no direct relaxation effect. Puerarin enhanced vasorelaxation to endothelium-independent relaxing agents, sodium nitroprusside and cromakalim. However, puerarin had no effect on vasorelaxation induced by endothelium-dependent relaxing agents, bradykinin and calcium ionophore A23187. The potentiating action of puerarin (10 microM) on sodium nitroprusside-mediated relaxation was not affected by the nitric oxide synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME; 300 microM), or by the disruption of the endothelium with Triton X-100. The effect of puerarin was reversible following a washout period. The potentiating effects were comparable with the 3'-5'-cyclic adenosine monophosphate (cyclic AMP) analogues, 8-bromoadenosine-3'-5'-cyclic monophosphate (8-Br-cyclic AMP; 10 muM) and Sp-isomer [S nomenclature refers to phosphorus] of adenosine-3', 5'-cyclic monophosphorothioate (Sp-cyclic AMPS; 3 microM), but not the 3'-5'-cyclic guanosine monophosphate (cyclic GMP) analogue, 8-bromoguanosine-3'-5'-cyclic monophosphate (8-Br-cyclic GMP; 3 microM). The cyclic AMP antagonist, Rp-isomer [R nomenclature refers to phosphorus] of 8-bromoadenosine-3', 5'-cyclic monophosphorothioate (Rp-8-Br-cyclic AMPS; 10 microM), but not cyclic GMP antagonist, Rp-isomer of 8-bromoguanosine-3', 5'-cyclic monophosphorothioate (Rp-8-Br-cyclic GMPS; 10 microM), reversed the effects of puerarin (10 microM) on the enhancement of vasorelaxation to sodium nitroprusside. Our results demonstrated that puerarin enhanced sodium nitroprusside-induced relaxation, possibly via the cyclic AMP-dependent pathway.

Publication Types: PMID: 17027964 [PubMed - indexed for MEDLINE]

81. Nitric oxide mediates the fungal elicitor-induced puerarin biosynthesis in Pueraria thomsonii Benth. suspension cells through a salicylic acid (SA)-dependent and a jasmonic acid (JA)-dependent signal pathway.

Yeung DK, Leung SW, Xu YC, Vanhoutte PM, Man RY.

Department of Biotechnology, Zhejiang Gongshang University, Hangzhou 310035, China. maojunxu@163.com

Nitric oxide (NO) has emerged as a key signaling molecule in plant secondary metabolite biosynthesis recently. In order to investigate the molecular basis of NO signaling in elicitor-induced secondary metabolite biosynthesis of plant cells, we determined the contents of NO, salicylic acid (SA), jasmonic acid (JA), and puerarin in Pueraria thomsonii Benth. suspension cells treated with the elicitors prepared from cell walls of Penicillium citrinum. The results showed that the fungal elicitor induced NO burst, SA accumulation and puerarin production of P. thomsonii Benth. cells. The elicitor-induced SA accumulation and puerarin production was suppressed by nitric oxide specific scavenger cPITO, indicating that NO was essential for elicitor-induced SA and puerarin biosynthesis in P. thomsonii Benth. cells. In transgenic NahG P. thomsonii Benth. cells, the fungal elicitor also induced puerarin biosynthesis, NO burst, and JA accumulation, though the SA biosynthesis was impaired. The elicitor-induced JA accumulation in transgenic cells was blocked by cPITO, which suggested that JA acted downstream of NO and its biosynthesis was controlled by NO. External application of NO via its donor sodium nitroprusside (SNP) enhanced puerarin biosynthesis in transgenic NahG P. thomsonii Benth. cells, and the NO-triggered puerarin biosynthesis was suppressed by JA inhibitors IBU and NDGA, which indicated that NO induced puerarin production through a JA-dependent signal pathway in the transgenic cells. Exogenous application of SA suppressed the elicitor-induced JA biosynthesis and reversed the inhibition of IBU and NDGA on elicitor-induced puerarin accumulation in transgenic cells, which indicated that SA inhibited JA biosynthesis in the cells and that SA might be used as a substitute for JA to mediate the elicitor- and NO-induced puerarin biosynthesis. It was, therefore, concluded that NO might mediate the elicitor-induced puerarin biosynthesis through SA- and JA-dependent signal pathways in wildtype P. thomsonii Benth. cells and transgenic NahG cells respectively.

Publication Types: PMID: 16989284 [PubMed - indexed for MEDLINE]

82. Modulation of high alcohol drinking in the inbred Fawn-Hooded (FH/Wjd) rat strain: implications for treatment.

Overstreet DH, Rezvani AH, Cowen M, Chen F, Lawrence AJ.

Bowles Center for Alcohol Studies & Department of Psychiatry, University of North Carolina at Chapel Hill, USA. dhover@med.unc.edu

The Fawn-Hooded rat (FH/Wjd) is an inbred alcohol-preferring rat strain, unlike most of the other strains that were selectively bred for high alcohol intake and preference. It was chosen for study some 16 years ago because of a reported mutation that disrupted platelet serotonin function. Although the FH/Wjd rat has high alcohol intake (>5 g/kg/day) and preference (>65%), interbreeding with an alcohol-non-preferring inbred strain suggested that these measures are unrelated to the serotonin abnormality. Similarly, the exaggerated immobility of the FH/Wjd rats in the forced swim test did not correlate with the high alcohol intake. Many compounds have been tested in the FH/Wjd rats after both acute and chronic treatment and a substantial number of them have proved effective. However, as the case with opiate antagonists, tolerance to the effects of the drug can develop. An up-regulation of opioid receptors accompanied the chronic treatment and this mechanism may account for the development of tolerance. Tolerance to opiate antagonists has also been demonstrated in two of the selectively bred alcohol-preferring rat lines, but it is unknown whether this process may contribute to the relapses seen in individuals being treated with naltrexone. Other drugs that reliably decrease alcohol intake in the FH/Wjd rats include the 5-hydroxytryptamine-2A receptor antagonist, amperozide, the mGlu5 receptor antagonist 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (MTEP) and herbal derivatives such as ibogaine, St. John's wort and kudzu extract. Thus, studies in the FH/Wjd rat have led to the discovery of a wide variety of targets for the development of novel agents to treat alcoholism. The fact that several of these drugs were shown to reduce alcohol intake in some of the selectively bred alcohol-preferring rat lines and/or alcohol-preferring vervet monkeys increases our confidence that they are good candidates for further development.

Publication Types: PMID: 16961764 [PubMed - indexed for MEDLINE]

83. One-pot synthesis of genistein from tyrosine by coincubation of genetically engineered Escherichia coli and Saccharomyces cerevisiae cells.

Katsuyama Y, Miyahisa I, Funa N, Horinouchi S.

Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan.

For production of genistein from N-acetylcysteamine-attached p-coumarate (p-coumaroyl-NAC) supplemented to the medium, a chalcone synthase (CHS) gene from Glycyrrhiza echinata, a chalcone isomerase (CHI) gene from Pueraria lobata, and an isoflavone synthase (IFS) gene from G. echinata were placed under the control of the galactose-inducible GAL promoters in pESC vector and were introduced in Saccharomyces cerevisiae. When the recombinant yeast cells (0.5 g wet weight) were used as "enzyme bags" and incubated at 30 degrees C for 48 h in 100 ml of the buffer containing galactose and 1 mM (265 mg/l) p-coumaroyl-NAC, ca. 340 microg genistein/l was produced. Another system consisting of two enzyme bags was also generated for the purpose of production of genistein from tyrosine. One enzyme bag was an Escherichia coli cell containing a phenylalanine ammonia-lyase gene from a yeast, a 4-coumarate/cinnamate:CoA ligase gene from the actinomycete Streptomyces coelicolor A3(2), the CHS gene, and the CHI gene, in addition to the acetyl-CoA carboxylase gene from Corynebacterium glutamicum, all of which were under the control of the isopropyl-beta-D-thiogalactopyranoside-inducible T7 promoter, and thus producing (S)-naringenin from tyrosine. The other enzyme bag was a S. cerevisiae cell containing the IFS gene. Coincubation of the E. coli cells (0.5 g wet weight) and S. cerevisiae cells (0.5 g wet weight) at 26 degrees C for 60 h in 20 ml of the buffer containing 3 mM (543 mg/l) tyrosine as the starting substrate yielded ca. 6 mg genistein/l.

Publication Types:PMID: 16960736 [PubMed - indexed for MEDLINE]

84. Puerariafuran, a new inhibitor of advanced glycation end products (AGEs) isolated from the roots of Pueraria lobata.

Jang DS, Kim JM, Lee YM, Kim YS, Kim JH, Kim JS.


Department of Herbal Pharmaceutical Development, Korea Institute of Oriental Medicine, 461-24 Jeonmin-dong, Yuseong-gu, Daejon 305-811, Korea.

A new 2-arylbenzofuran, puerariafuran (1), as well as three known compounds, coumestrol (2), daidzein (3), and genistein (4), were isolated from a MeOH extract of the roots of Pueraria lobata as active constituents, using an in vitro bioassay based on the inhibition of advanced glycation end products (AGEs) to monitor chromatographic fractionation. The structure of 1 was determined by spectroscopic data interpretation, particularly by extensive 1D and 2D NMR studies. All the isolates (1-4) were evaluated for the inhibitory activity on AGEs formation in vitro.

Publication Types: PMID: 16946543 [PubMed - indexed for MEDLINE]

85. Major isoflavonoid contents of the phytoestrogen rich-herb Pueraria mirifica in comparison with Pueraria lobata.

Cherdshewasart W, Subtang S, Dahlan W.

Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Patumwan, Bangkok 10330, Thailand. cwichai@sc.chula.ac.th

Pueraria mirifica tubers collected from 28 out of 76 provinces of Thailand and Pueraria lobata tubers collected from Guangzhou province, China were submitted to HPLC analysis with the established gradient system comprising 1.5% acetic acid and acetonitrile. Five major isoflavonoids, including puerarin, daidzin, genistin, daidzein and genistein, were adopted as authentic standards. P. mirifica tubers showed intra- as well as inter-provincial differences in isoflavonoid and total isoflavonoid contents. The difference in both cases should be mostly influenced by genetic and environmental factors. In comparison with P. lobata, P. mirifica population exhibited differences only with a lower amount of daidzein.

Publication Types: PMID: 16930918 [PubMed - indexed for MEDLINE]

86. Evaluation of oriental medicinal herbs for estrogenic and antiproliferative activities.

Kang SC, Lee CM, Choi H, Lee JH, Oh JS, Kwak JH, Zee OP.

College of Pharmacy, SungKyunKwan University, Suwon, Gyonggi-Do, South Korea.

Herb extracts commercially used in Asia were screened for their estrogenic activity with a recombinant yeast system with both a human estrogen receptor (ER) expression plasmid and a reporter plasmid. Pueraria lobata (flower) had the highest estrogenic relative potency (RP, 17-estradiol = 1.00) (7.8e-3) (RP for + control), followed by Amomum xanthioides (1.3e-3), Glycyrrhiza uralensis, Zingiber officinale, Rheum palmatum, Curcuma aromatica, Eriobotrya japonica, Sophora flavescens, Anemarrhena asphodeloides, Polygonum multiflorum and Pueraria lobata (root) (9.5e-4-1.0e-4), and Prunus persica, Lycoppus lucidus and Adenophora stricta (9.0e-5-8.0e-5). In the antiproliferative assay, five human cancer cell lines representing different tissues (breast, lung and ovary) were used. Eriobotrya japonica showed strong cytotoxicity in ER-negative breast cancer (MDA-MB-231), cervix epitheloid (HeLa) and lung (A549) carcinoma cell lines. Copyright (c) 2006 John Wiley & Sons, Ltd.

Publication Types: PMID: 16906642 [PubMed - indexed for MEDLINE]

87. Pharmacokinetic profile of the isoflavone puerarin after acute and repeated administration of a novel kudzu extract to human volunteers.

Penetar DM, Teter CJ, Ma Z, Tracy M, Lee DY, Lukas SE.

Behavioral Psychopharmacology Research Laboratory, McLean Hospital, Belmont, MA 02478, USA. dpenetar@mclean.harvard.edu

OBJECTIVES: This study was undertaken to assess the pharmacokinetic profile of puerarin, the major isoflavone found in a kudzu (Pueraria lobata) extract after acute and repeated administration. METHODS: Participants were given either single or repeated doses of kudzu extract, and blood samples were collected for either 8 or 72 hours for subsequent pharmacokinetic analyses of puerarin. RESULTS: Using WinNonlin pharmacokinetic data analysis software, puerarin was found to be rapidly absorbed via the oral route, reach peak levels at 2 hours, and have a half-life of approximately 4.3 hours. The elimination half-life was not significantly altered after repeated administration. CONCLUSIONS: A formulation of kudzu extract delivers a large amount of the principal isoflavone in a rapid manner. The elimination rate constants and the mono-exponential decline in blood levels suggest that a one compartment model adequately explains how puerarin is handled by the body. Three times a day dosing is recommended as accumulation will not occur, and plasma levels remain at levels that are biologically active, even 8 hours after the last steady-state dose.

Publication Types: PMID: 16884345 [PubMed - indexed for MEDLINE]

88. [Amebicidal plants extracts]

[Article in Polish]

Derda M, Hadaś E, Thiem B, Sułek A.

Katedra i Zakład Biologii i Parazytologii Lekarskiej, Akademia Medyczna, ul. Fredry 10, 61-701 Poznań.

The free-living amoebae from genus Acanthamoeba are the causative agents of granulomatous amebic encephalitis (GAE), a chronic progressive disease of the central nervous system; amebic keratitis (AK), a chronic eye infection; amebic pneumitis (AP), a chronic lung infection, and skin infection. Chemotherapy of Acanthamoeba infection is problematic. The majority of infections have been fatal. Only a few cases are reported to have been treated successfully with very highly toxic drugs. The therapy might be succeed, if the diagnosis and therapy is made at very early stage of infection. In our experiments we used the following plant extracts: Solidago virgaurea, Solidago graminifolia, Rubus chamaemorus, Pueraria lobata, and natural plants products as ellagic acid and puerarin. Those therapeutic agents and plants extracts have been tested in vitro for amebicidal or amebostatic activity against pathogenic Acanthamoeba spp. Our results showed that methanol extracts obtained from plants are active against axenic pathogenic Acanthamoeba sp. trophozoites in vitro at concentration below 0.1 mg/ml. Further studies are needed to investigate whether these extracts are also effective in vivo in animal model of infection with Acanthamoeba sp.

Publication Types: PMID: 16862806 [PubMed - indexed for MEDLINE]

89. Effects of Pueraria mirifica on vascular function of ovariectomized rabbits.

Wattanapitayakul SK, Chularojmontri L, Srichirat S.

Department of Pharmacology, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Wattana, Bangkok 10110, Thailand. suvara@swu.ac.th

Estrogen stimulates endothelial nitric oxide (NO) production and attenuates endothelial dysfunction in ischemia/repurfusion and menopause. Recent studies have shown that phytoestrogens from dietary sources improve endothelial function and reduce cardiovascular risks. The Thai medicinal plant Pueraria mirifica (PM) contains many potent phytoestrogens including miroestrol and deoxymiroestrol but no study on vascular function has been established. Ground powder of PM was orally given to ovariectomized White New Zealand rabbits (OVX + PM group) (n = 4) weighing 3.2-4.0 kg at the dose of 100 mg/kg for 90 days. Saline-treated ovariectomized rabbits were assigned as a control group (OVX group) (n = 5). At the end of treatment thoracic aorta was isolated for functional evaluation. Maximal relaxant response to acetylcholine (ACh) was significantly increased (24%) with 3.5-fold decrease in EC50 while no change in relaxant response to sodium nitroprusside was observed Minimal and maximal responses to 17beta-estradiol (E2) were increased in the OVX + PM group and L-NAME (100 mM) attenuated Emax of E2. PM significantly decreased maximal contractile responses to norepinephrine (NE), but no change in EC50 was observed. In addition to vascular study, the authors found no significant alteration in serum cholesterol, LDL, triglyceride, HDL, ALT AST alkaline phosphatase, and lipid peroxidation in OVX + PM rabbits. These data demonstrate that PM (100 mg/kg/d) improved endothelial function through NO-dependent pathway and increased response to E2 while sensitivity to NE was reduced. In addition, it had no impact on lipid profile, liver enzymes, and ALP activities. PM is a potential source of phytoestrogens for postmenopausal women to improve cardiovascular function or reduce cardiovascular risks.

Publication Types: PMID: 16862667 [PubMed - indexed for MEDLINE]

90. Production of puerarin and isoflavones in cell suspension cultures of Pueraria lobata (Willd.): effects of medium supplementation with casein hydrolysate and coconut milk.

Li L, Zhang CR.

College of Life Science, South China Normal University, Guangzhou, Guangdong-510631, China. liling@scnu.edu.cn

Callus induced from leaf explants of Pueraria lobata seedlings were suspended in Gamborg B5 medium supplemented with 1 mg l(-1) 2,4-dichlorophenoxyacetic acid, 1 mg l(-1) naphthalene acetic acid, 0.5 mg l(-1) kinetin and 30 g l(-1) sucrose. The effects of coconut milk and casein hydrolysate (CH) on cell growth and yields of puerarin and isoflavones in cells suspension were studied. The contents of total isoflavones and puerarin in suspension cultures were determined by spectrophotometry and HPLC. Coconut milk (10%, filter sterilized) decreased the growth of cell cultures and the accumulation of total isoflavones, while 0.2% CH promoted the growth of cell cultures and the accumulation and release of puerarin and total isoflavones. The total yield of puerarin and isoflavones were 34% and 40.8% higher than in the control, respectively. The optimum medium for cell cultures of leaves of P. lobata seedlings was B5 liquid medium supplemented with 2% sucrose, 1.0 mg l(-1) 2,4-D, 1.0 mg l(-1) NAA, 0.5 mg l(-1) kinetin and 20 mg l(-1) CH. The procedure use is a potentially useful for the production of isoflavones.

Publication Types: PMID: 16850870 [PubMed - indexed for MEDLINE]

91. [The experimental study of Radix Puerariae inhibiting glycation in rats induced by D-galactose]

[Article in Chinese]

Zhang SP, Fang WJ, Lü JH, Tan HR, Pan JQ.

Department of Pharmacology, Pharmacy College of Jinan University, Guangzhou 510632, China.

OBJECTIVE: To observe the effects of Radix Puerariae on protein glycation in model rats induced by D-galactose. METHODS: The model rats of protein glycatin were induced by intraperitoneal administration of D-galactose (150 mg/kg) for 8 weeks, and all rats were treated by Radix Puerariae (High dose 300 mg/kg, Middle dose 150 mg/kg, Low dose 75 mg/kg) for 6 weeks. The activity of aldose reductase in red blood cells, the content of fructosamine in serum, the amount of glycohaemoglobin and advanced glycation end-products, the level of insulin in serum, the activity of superoxide dismutase and the amount of maleic dialdehyde were measured. RESULTS: High dose and middle dose of Radix Puerariae could decrease the level of blood glucose and the activity of aldose reductase in red blood cells, inhibit the formation of glycation products significantly in model rats induced by D-galactose (P < 0.01), increase insulin sensitivity and activity of superoxide dismutase (P < 0.01) and decrease the amount of maleic dialdehyde (P < 0.01). CONCLUSION: Puerariae can significantly inhibit glycation reaction in rats induced by D-galactose.

Publication Types: PMID: 16850728 [PubMed - indexed for MEDLINE]

92. Regulation of isoflavone production in hydroponically grown Pueraria montana (kudzu) by cork pieces, XAD-4, and methyl jasmonate.

Kirakosyan A, Kaufman PB, Chang SC, Warber S, Bolling S, Vardapetyan H.

Department of Cardiac Surgery, University of Michigan, Ann Arbor, MI 48109-0686, USA. pbk@umich.edu

A mini-hydroponic growing system was employed for seedlings of kudzu vine (Pueraria montana) and contents of isoflavones (daidzein, genistein, daidzin, genistin, and puerarin) from shoot and root parts of seedlings were analyzed quantitatively. In addition, exogenous cork pieces, polymeric adsorbent, XAD-4, and universal elicitor, methyl jasmonate (MeJA), were used to regulate the production of these isoflavones. It was shown that cork pieces up-regulate the production of daidzein and genistein up to seven- and eight-fold greater than the levels obtained for control roots. In contrast, levels of glucosyl conjugates, daidzin and genistin, decrease up to five- and eight-fold, respectively. Cork treatment also induces the excretion of the root isoflavone constituents into the growth medium. Minimal levels of isoflavones are absorbed by the cork pieces. XAD-4 stimulates the production of glucosyl conjugates, daidzin and genistin, in root parts about 1.5-fold greater than that obtained in control roots. These are the highest amounts of daidzin and genistin that are observed (5.101 and 6.759 mg g(-1) dry weight, respectively). In contrast to these two adsorbents, MeJA increases the accumulation of isoflavones in shoot rather than in root parts of seedlings, about three- to four-fold over control levels, with the exception of genistein. These studies reveal new observations on the regulation of isoflavone production in hydroponically grown Pueraria montana plants by two adsorbents (cork pieces and XAD-4) and MeJA elicitor.

Publication Types: PMID: 16841218 [PubMed - indexed for MEDLINE]

93. [Experimental study of Bailian Caogen granule on pharmacodynamics]

[Article in Chinese]

Zhao RY, Zhao LN, Zhou YX, Li DM, Hao W, Chen KM.

Dept of Pharmacology, Shenyang Medical College, China.

OBJECTIVE: To study the analgesic, antipyretic and anti-inflammatory effect of Bailian Caogen granule. METHOD: The antipyretic effects of Bailian Caogen granule was evaluated in rabbit fever model induced by peptone. The analgesic effect of the drug was studied with pain model of mice induced by acetic acid and hot plate, The severity of oedema in inflamed animal was observed to study the anti-inflammatory effects of Bailian Caogen granule. RESULT: Bailian Caogen granule could obviously inhibit the fever of rabbits. The number of writhing induced by acetic acid was reduced and the pain threshold of mice was increased by Bailian Caogen granule. Bailian Caogen granule also had anti-inflammatory activity against xylene-induced mouse ear swelling and carrageenin-induced rat paw edema. CONCLUSION: Bailian Caogen granule has significant analgesic, antipyretic and anti-inflammatory activities.

Publication Types: PMID: 16830829 [PubMed - indexed for MEDLINE]

94. KIOM-79 inhibits LPS-induced iNOS gene expression by blocking NF-kappaB/Rel and p38 kinase activation in murine macrophages.

Jeon YJ, Li MH, Lee KY, Kim JS, You HJ, Lee SK, Sohn HM, Choi SJ, Koh JW, Chang IY.

College of Medicine, Chosun University, 375 Susukdong, Kwangju 501-709, Republic of Korea.

We demonstrate that KIOM-79, combined extracts obtained from Magnolia officinalis, Pueraria lobata, Glycyrrhiza uralensis, and Euphorbia pekinensis, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Treatment of RAW 264.7 cells with KIOM-79 inhibited LPS-stimulated nitric oxide production in a dose-related manner. Immunohisto-chemical staining of iNOS and RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression. Immunostaining of p65, EMSA, and reporter gene assay showed that KIOM-79 inhibited NF-kappa/Rel nuclear translocation, DNA binding, and transcriptional activation, respectively. Western immunoblot analysis of p38 kinase showed KIOM-79 significantly inhibited the phosphoylation of p38 kinase which is important in the regulation of iNOS gene expression. Collectively, this series of experiments indicates that KIOM inhibits iNOS gene expression by blocking NF-kappa/Rel and p38 kinase signaling. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of KIOM-79 on iNOS suggest that KIOM-79 may represent a useful anti-inflammatory agent.

Publication Types: PMID: 16806764 [PubMed - indexed for MEDLINE]

95. Effect of Pueraria mirifica on the sexual skin coloration of aged menopausal cynomolgus monkeys.

Trisomboon H, Malaivijitnond S, Cherdshewasart W, Watanabe G, Taya K.

Department of Physiology, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.

To investigate the estrogenic effect of Pueraria mirifica (PM), a Thai herbal plant that contains many phytoestrogens, sexual skin coloration was studied in cynomolgus monkeys. Aged menopausal monkeys were divided into three groups. Each group (n=3) was fed 10, 100, or 1,000 mg of PM daily. The treatment schedule was divided into three periods, a 30-day pre-treatment period, 90-day treatment period, and 60-day post-treatment period. The results show that the sexual skin exhibited reddish coloration within 24 h after PM-treatment and remained this way for the first half of the PM-feeding period. The changes in sexual skin coloration were not dose-dependent. The present results indicate that PM had estrogenic action by increasing reddish sexual skin coloration in aged menopausal monkeys.

Publication Types: PMID: 16799265 [PubMed - indexed for MEDLINE]

96. Both soybean and kudzu phytoestrogens modify favorably the blood lipoprotein profile in ovariectomized and castrated hamsters.

Guan L, Yeung SY, Huang Y, Chen ZY.

Li Ka Shing Institute of Health Sciences, Food and Nutritional Sciences Program of the Department of Biochemistry, and Department of Physiology, The Chinese University of Hong Kong, Shatin, NT, Hong Kong, China.

The present study compared the hypolipidemic activity of kudzu phytoestrogens with that of soybean phytoestrogen in estrogen- and androgen-deficient hamsters. In the first experiment, ovariectomized hamsters (n = 37) were randomly divided into four groups (n = 9-10 each group). The first group was the control group, whereas the second group had the time-releasing estradiol-17beta subcutaneous (pellet) implants as a positive control. The third and fourth groups were orally administered soybean or kudzu phytoestrogen extracts (30 mg/kg of body weight) per day. In the second experiments, the first group of male hamsters (n = 9) received a sham operation, whereas the other three groups of male hamsters (n = 9 each) were castrated. The castrated control group received orally distilled water, whereas the second and third castrated groups were orally given 30 mg/kg soybean or kudzu phytoestrogen extracts. The results for the first experiment showed that the ovariectomized hamsters orally given soybean and kudzu phytoestrogen extracts had significantly decreased serum total cholesterol (TC) and non-high-density lipoprotein cholesterol (non-HDL-C) with HDL cholesterol (HDL-C) being unaffected. The data from the second experiment demonstrated that administration of soybean but not kudzu phytoestrogen extracts decreased significantly serum TC. However, administration of kudzu phytoestrogens caused redistribution of cholesterol among lipoproteins, leading to a significant decrease in the ratio of non-HDL-C to HDL-C. It was concluded that both soybean and kudzu phytoestrogens could modify favorably lipoprotein profiles in ovariectomized and castrated hamsters.

PMID: 16787047 [PubMed - indexed for MEDLINE]

97. [Protective effect of pueraria flavonoid on the cerebral ischemic reperfusion injury in rats]

[Article in Chinese]

Wang PY, Wang HP, Li GW.

Department of Neurobiology, Anhui Medical University, Hefei 230032, China.

OBJECTIVE: To research the protective effect of pueraria flavonoid on the cerebral ischemic reperfusion injury. METHOD: Using the middle cerebral artery occlusion model (MCAO) in rats, we investigated the influence of pueraria flavonoid on the brain water content, the infarct volume, the activities of SOD, and the content of MDA. RESULT: Pueraria Flavonoid could obviously reduce the brain water content and the infract volume in MCAO, increase the activities of SOD, and decrease the content of MDA in the cerebral ischemia- reinfusion model of rats. CONCLUSION: Pueraria has the function of scavenging free radicals and the protective effect on ischemic brain tissue.

Publication Types: PMID: 16780163 [PubMed - indexed for MEDLINE]

98. Management of SAH with traditional Chinese medicine in China.

Wang C, Zhao X, Mao S, Wang Y, Cui X, Pu Y.

Department of Neurology, Beijing Tiantan Hospital, affiliated to the Capital University of Medical Sciences, Beijing, China. snow_sen@yahoo.com

China lacks large scale authorized epidemiological study results in allusion to subarachnoid hemorrhage (SAH) within recent 15 years since MONICA (multinational monitoring of trends and determinants in cardiovascular disease) study revealed SAH situation in China in 2000. The main cause of SAH in China is aneurysm which takes up 30-50%, while over 90% aneurysm locates at Willis circle. Early surgery for SAH after aneurysm rupture is the dominant procedure to deal with SAH in China. Moreover, calcium antagonists rank the absolute leading position for cerebral vascular spasm (CVS) among medication-based treatment options. However, traditional Chinese medicine such as Salvia miltiorrhiza, Acanthopanax senticosus, Ginkgo biloba, Pueraria lobata, Liguisticum chuanxiong, cow bezoar, Diospyros kaki and Gynostemma pentaphyllum have been proven beneficial in CVS prevention and treatment, while Salvia miltiorrhiza and TCM soup have unique effects on bleeding absorption. In addition, aescine and some TCM soup might relieve strong headache after SAH. In general, TCM integrated with western medicine have shown unique advantages in the current treatment of SAH in China. However, it is a pity that China still lacks larger scale randomized controlled trials and research on SAH treatment focusing on TCM and the related mechanism of TCM on SAH still need to be investigated further.

Publication Types: PMID: 16759447 [PubMed - indexed for MEDLINE]

99. Estrogenic effect of main components kakkalide and tectoridin of puerariae flos and their metabolites.

Shin JE, Bae EA, Lee YC, Ma JY, Kim DH.

College of Pharmacy, Kyung Hee University.

To understand the relationship between the metabolism and estrogenic activity of kakkalide and tectoridin, main isoflavones in the flower of Pueraria thunbergiana (family Leguminosae), these isoflavones and their metabolites by human intestinal microflora as well as their estrogenic effects were investigated. All human fecal specimens metabolized kakkalide and tectoridin. All isolated kakkalide-hydrolyzing intestinal bacteria also hydrolyzed kakkalide and tectoridin to irisolidone and tectorigenin, respectively. When the estrogenic effects of kakkalide and tectoridin were compared with those of their metabolites irisolidone and tectorigenin, the metabolites more potently increased proliferation of MCF-7 cells than kakkalide and tectoridin. These metabolites also potently induced estrogen-response c-fos and pS2 mRNA expression. These results suggest that kakkalide and tectoridin may be metabolized mainly to irisolidone and tectorigenin, respectively, by intestinal microflora in the intestines, and which may be subsequently absorbed into the blood where they can express their estrogenic effect.

PMID: 16755017 [PubMed - in process]

101. Separation and determination of isoflavonoids in several kudzu samples by high-performance capillary electrophoresis (HPCE).

Fang C, Wan X, Tan H, Jiang C.

Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, Anhui, China.

Pueraria lobata is a rich source of isoflavonoids. The detection and identification of isoflavonoid components from root, stem, leaf, callus and cell samples, is very important for the best, safest and most efficacious use of kudzu as a medicinal plant, and for the studies on quantitative analysis in the secondary metabolism of isoflavonoids. In this paper, a simple, rapid and precise high-performance capillary electrophoresis (HPCE) method with diode array detection (DAD) has been developed for separation and determination of isoflavonoids in several kudzu samples. The isoflavonoids could be well separated within 15 min in a 40 cm length capillary at a separation voltage of 15kV in a 30 mmol L(-1) borax buffer (pH9.29), and this proposed method demonstrated excellent reproducibility and accuracy with relative standard deviations of less than 5% for isoflavonoid content (n = 5) of different kudzu samples. The relationship between peak areas and isoflavone concentrations, in a specified working range with linear response, was determined by first-order polynomial regression over the range 0.05-0.5 mg mL(-1) for puerarin and 2.5-50 microg mL(-1) for 3'-methoxypuerarin, daidzin and daidzein, respectively, and quantitative evaluation of those four main isoflavonoid components was determined by ultraviolet absorption at lambda = 192 nm. The differences were also illustrated by comparison of the determination of isoflavonoid components from kudzu root, stem, leaf samples and plant tissue cultures in vitro.

Publication Types: PMID: 16734027 [PubMed - indexed for MEDLINE]

102. Using vaginal cytology to assess the estrogenic activity of phytoestrogen-rich herb.

Malaivijitnond S, Chansri K, Kijkuokul P, Urasopon N, Cherdshewasart W.

Primate Research Unit, Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Patumwan, Bangkok 10330, Thailand. Suchinda.m@chula.ac.th

To assess the estrogenic activities of synthetic estrogen, synthetic phytoestrogen, Pueraria lobata and three distinct cultivars of Pueraria mirifica, a phytoestrogen-rich herb, a vaginal cytology assay in ovariectomized rats were used. Rats were ovariectomized and treated with DW, estradiol valerate (1 mg/kg BW), genistein (0.25-2.5 mg/kg BW), Pueraria lobata and Pueraria mirifica (10-1,000 mg/kg BW) for 14 days. The vaginal cytology was checked daily and the uteri were dissected and weighed at the end of treatment or post-treatment periods. The treatments of DW, genistein and Pueraria lobata did not influence the vaginal epithelium, but the injection of estradiol valerate induced a vaginal cornification from day-3 of treatment to day-14 of post-treatment period. The occurrence of vaginal cornification after treatment and the recovery after the cessation was dependent on dosages and cultivars of Pueraria mirifica. The increments of uterus weight in all rats agreed with the cornification of vaginal epithelium. Although both uterotropic and vaginal cytology assays can be used to assess the estrogenic activity of phytoestrogen-rich herb, however, using vaginal cytology assay has two advantages: (1) we do not need to kill the animals and (2) we can follow up the recovery after the cessation of treatment.

Publication Types: PMID: 16730147 [PubMed - indexed for MEDLINE]

103. [Protective effect of puerarin on stress-induced gastric mucosal injury in rats]

[Article in Chinese]

Wang FW, Li J, Hu ZL, Xie YY.

Institute of Materia Medica, Shandong Academy of Medical Sciences, Jinan 250062, China. wangfuwww@tom.com

OBJECTIVE: To study the protective effect of puerarin on stress-induced gastric mucosal injury in rats. METHOD: The model of gastric ulcer was established by restraint plus water-immersion stress in rats. Gastric motility was monitored by the method of "Gas Balloon". Gastric mucosal blood flow was recorded by laser-Doppler flowmetry. Colorimetric method was used to determine the content of NO and ET in gastric mucosal tissue. Meantime the pathologic changes of gastric mucosal was examined. RESULT: Puerarin could significantly attenuated gastric mucosal damage induced by water-immersion stress, inhibited gastric motility, specially decreased the index of gastric motility and percentage of gastric contraction time and numbers of violent contraction. The gastric mucosal blood flow and NO level in gastric mucosal were enhanced, while ET level was reduced by puerarin. The degree of tissue damage in gastric mucosal was also significantly attenuated after administration fo puerarin. CONCLUSION: Puerarin exerts a significant protective effect on water-immersion stress-induced gastric mucosal damage by relaxing the vessels, increasing NO level in gastric mucosal, increasing regional gastric mucosal blood flow and inhibiting gastric motility.

Publication Types: PMID: 16722386 [PubMed - indexed for MEDLINE]

104. [Puerarin suppresses the proliferation of vscular smooth muscule cells and c-fos and bcl-2 protein expression]

[Article in Chinese]

Xu YZ, Gao Y, Li PZ, Wang NF, Xu HY, Tong GX.


The First People's Hospital of Hangzhou, Hangzhou 310006, China. qqyzxu@zj165.com

OBJECTIVE: To observe the role of puerarin on the proliferation of vascular smooth muscle cells(VSMC) induced by thrombin (T) and the effect of puerarin on the c-fos and bcl-2 protein expression. METHOD: Cell number and cell cycle analysis using flow cytometry were adopted as two different indicators of effects on proliferation of VSMC. Western blot was used to indicate the changes of c-fos and bcl-2 protein after 24 h of treatment of T and puerarin. RESULT: 1.5 x 10(-5) - 1.5 x 10(-3) mol x L(-1) puerarin could significantly suppress this stimulation of VSMC proliferation and DNA synthesis induced by T. Western blot demonstrated that after 24 hour of treatment with T and puerarin, T could significantly increase c-fos and bcl-2 protein and 1.5 x 10(-5) - 1.5 x 10(-3) mol x L(-1) puerain could significantly suppress this increase. CONCLUSION: puerarin can suppress the proliferation and DNA synthesis of VSMC promoted by T. This inhibitory effects of puerarin are closely related with the suppression of c-fos and bcl-2 protein.

Publication Types: PMID: 16722382 [PubMed - indexed for MEDLINE]

105. High-performance thin-layer chromatographic fingerprints of isoflavonoids for distinguishing between Radix Puerariae Lobate and Radix Puerariae Thomsonii.

Chen SB, Liu HP, Tian RT, Yang DJ, Chen SL, Xu HX, Chan AS, Xie PS.

State Key Laboratory of Chinese Medicine and Molecular Pharmacology, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Hong Kong, China.

The roots of Pueraria lobata (Wild.) Ohwi and Pueraria thomsonii Benth have been officially recorded in all editions of Chinese Pharmacopoeia under the same monograph 'Gegen' (Radix Puerariae, RP). However, in its 2005 edition, the two species were separated into both individual monographs, namely 'Gegen' (Radix Puerariae Lobatae, RPL) and 'Fenge' (Radix Puerariae Thomsonii, RPT), respectively, due to their obvious content discrepancy of puerarin, the major active constituent. In present paper, the fingerprint of high-performance thin-layer chromatography (HPTLC) combining digital scanning profiling was developed to identify and distinguish the both species in detail. The unique properties of the HPTLC fingerprints were validated by analyzing ten batches of Pueraria lobata and P. thomsonii samples, respectively. The common pattern of the HPTLC images of the roots of Pueraria spp. and the respective different ratios of the chemical distribution can directly discern the two species. The corresponding digital scanning profiles provided an easy way for quantifiable comparison among the samples. Obvious difference in ingredient content and HPTLC patterns of the two species questioned their bio-equivalence and explained that recording both species separately in the current edition of Chinese Pharmacopoeia (2005 edition) is reasonable due to not only the content of major constituent, puerarin, but also the peak-to-peak distribution in the fingerprint and integration value of the total components. Furthermore, the HPTLC fingerprint is also suitable for rapid and simple authentication and comparison of the subtle difference among samples with identical plant resource but different geographic locations.

Publication Types: PMID: 16714027 [PubMed - indexed for MEDLINE]

105. [Protective effects of hydroxyethylpuerarin against brain astrocytes injury induced by hydrogen peroxide]

[Article in Chinese]

Zhang B, Wei XB, Liu HQ, Wang LX, Sun R, Zhang XM.

Institute of Pharmacology, School of Medicine, Shandong University, Jinan 250012, China.

AIM: To study the protective effects of hydroxyethylpuerarin against the injury of astrocytes induced by hydrogen peroxide (H2O2). METHODS: Experiments were performed with cells from passage 4. Plasma membrane integrity was measured by lactate dehydrogenase (LDH) release. The occurrence of apoptosis was measured by flow cytometry. The glutamate uptake of astrocytes was studied with [3H]-glutamate incorporation. Intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were assessed by automatic biochemistry analyzer. RESULTS: Compared with H2O2 injured group, the occurrence of apoptosis, levels of LDH release and intracellular MDA of astrocytes reduced in hydroxyethylpuerarin pre-treated groups, but the glutamate uptake and intracellular SOD activity of astrocytes increased. CONCLUSION: Hydroxyethylpuerarin could reduce the occurrence of apoptosis and improve neurotrophic function of astrocytes, which may be related with its antioxidant effects during oxidative stress.

Publication Types: PMID: 16671550 [PubMed - indexed for MEDLINE]

107. The estrogenic effect of Pueraria mirifica on gonadotrophin levels in aged monkeys.

Trisomboon H, Malaivijitnond S, Watanabe G, Cherdshewasart W, Taya K.

Department of Physiology, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand.

We investigated the effect of Pueraria mirifica (PM) on gonadotrophin and estradiol levels in aged animals; nine menopausal cynomolgus monkeys were divided into three groups. Each group (n = 3) was fed with 10, 100, and 1000 mg/d of PM for 90 d. PM-10 induced the decrease of follicle stimulating hormone (FSH) levels on d 15-90 in one out of three monkeys. PM-100 and PM-1000 decreased FSH levels of all monkeys throughout the treatment period. After the treatment period, FSH levels continued to decrease for 5 and 10-20 d in PM-100 and PM-1000, respectively, and the levels rebounded in all groups thereafter. PM-10 decreased luteinizing hormone (LH) levels throughout the treatment period in one out of three monkeys and returned to the pretreatment levels immediately after stopping treatment. PM-100 and PM-1000 prominently decreased LH levels between d 10 and 90 during treatment and persisted until d 15-25 and d 20-30 for PM-100 and PM-1000, respectively, during the post-treatment period. Serum LH levels rebounded after returning to pre-treatment levels in a dose-dependent manner. Estradiol levels tended to decrease during the treatment period in all groups. The daily feeding of PM suppressed gonadotrophin levels in aged menopausal monkeys based on dose. Moreover, they can be recovered, and there is a direct correlation between dosage and recovery time. PM may be effective as an alternative medicine in menopausal women because the effects are not permanent.

Publication Types: PMID: 16622301 [PubMed - indexed for MEDLINE]

108. Identification of isoflavonoids in several kudzu samples by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

Fang C, Wan X, Tan H, Jiang C.

Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and Ministry of Agriculture, P.R. China.

Pueraria lobata is a rich source of isoflavonoids. The detection and identification of isoflavonoid components from Pueraria radix (RP), callus and cell cultures, is very important for the safest and most effective use of kudzu as a medicinal plant, and for the studies on quantitative analysis and secondary metabolism of isoflavonoids in vitro cultures. Liquid chromatography is coupled with negative and positive electrospray ionization (ESI) tandem mass spectrometry (MS-MS), and photodiode array detection is used to characterize and detect isoflavonoids in root, callus, and cell samples of P. lobata. Characteristic product ions of aglycones, O-glucosides, and C-glucosides were obtained from the full-scan ESI-MS chromatography of the major peaks and the MS-MS spectra of the protonated ions. Five major components of puerarin, daidzin-6"-O-acetylester, genistin-6"-O-malonylester, biochanin A-7-O-glucoside-6"-O-malonylester, and daidzein are detected and identified from the methanolic extract of P. lobata callus cultures. The major isoflavonoid components of P. lobata cell suspension cultures are identified as puerarin, daidzin, daidzin-6"-O-acetylester, genistin-6"-O-malonylester, biochanin A-7-O-glucoside-6"-O-malonylester, genistein-8-C-glucoside-6"-O-malonylester, and daidzein, on the basis of ESI-MS and MS-MS spectra analysis. Likewise, puerarin, daidzin, genistein-6"-O-malonylester, 3'-methoxypuerarin, and daidzein are detected and identified from RP. Of those isoflavonoid components detected, daidzin-6"-O-acetylester is a new isoflavonoid glucoside and is for the first time detected from P. lobata cultures in vitro.

Publication Types: PMID: 16620495 [PubMed - indexed for MEDLINE]

109. Differential effects of isoflavones, from Astragalus membranaceus and Pueraria thomsonii, on the activation of PPARalpha, PPARgamma, and adipocyte differentiation in vitro.

Shen P, Liu MH, Ng TY, Chan YH, Yong EL.

Department of Obstetrics and Gynecology, Yong Loo Lin School of Medicine, National University of Singapore, 119074, Singapore.

Compounds that target the peroxisome proliferator-activated receptors PPARalpha and PPARgamma are used to correct dyslipidemia and to restore glycemic balance, respectively. Because the majority of diabetic patients suffer from atherogenic lipid abnormalities, in addition to insulin resistance, ligands are required that can activate both PPARalpha and PPARgamma. In this study, we used chimeric PPARalpha/gamma reporter-gene bioassays to screen herbal extracts with purported antidiabetic properties. Extracts of Astragalus membranaceus and Pueraria thomsonii significantly activated PPARalpha and PPARgamma. Bioassay-guided fractionation resulted in the isolation of the isoflavones, formononetin, and calycosin from Astragalus membranaceus, and daidzein from Pueraria thomsonii as the PPAR-activating compounds. We investigated the effects of these and 2 common isoflavones, genistein and biochanin A, using chimeric and full-length PPAR constructs in vitro. Biochanin A and formononectin were potent activators of both PPAR receptors (EC50 = 1-4 micromol/L) with PPARalpha/PPARgamma activity ratios of 1:3 in the chimeric and almost 1:1 in the full-length assay, comparable to those observed for synthetic dual PPAR-activating compounds under pharmaceutical development. There was a subtle hierarchy of PPARalpha/gamma activities, indicating that biochanin A, formononetin, and genistein were more potent than calycosin and daidzein in chimeric as well as full-length receptor assays. At low doses, only biochanin A and formononetin, but not genistein, calycosin, or daidzein, activated PPARgamma-driven reporter-gene activity and induced differentiation of 3T3-L1 preadipocytes. Our data suggest the potential value of isoflavones, especially biochanin A and their parent botanicals, as antidiabetic agents and for use in regulating lipid metabolism.

Publication Types: PMID: 16549448 [PubMed - indexed for MEDLINE]

110.Easy replication of Pueraria Lobata toward hierarchically ordered porous gamma-Al2O3.

Li C, He J.

Functional Nanomaterials Laboratory, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences (CAS), Chaoyangqu Datunlu Jia 3, Beijing 100101, China.

Hierarchically ordered porous alumina was prepared via a facile immersion-fuming-calcination process using Pueraria lobata as template. The as-prepared alumina inherited nearly all morphological features of the template, as shown by SEM observations. It also contains abundant mesopores based on nitrogen adsorption-desorption measurements. The crystalline phase of the as-prepared alumina was ascertained to be gamma-alumina by analyzing its XRD pattern. Pt nanoparticles were in situ synthesized in the gamma-alumina matrix and annealed at different temperatures in N(2) atmosphere. TEM observations showed that Pt nanoparticles supported by the as-prepared alumina have significantly high thermal stability.

PMID: 16519490 [PubMed]

111. Puerarin decreases serum total cholesterol and enhances thoracic aorta endothelial nitric oxide synthase expression in diet-induced hypercholesterolemic rats.

Yan LP, Chan SW, Chan AS, Chen SL, Ma XJ, Xu HX.

Institute of Medicinal Plant Development, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.

Hypercholesterolemia is a dominant risk factor for the development and progression of atherosclerosis and cardiovascular diseases. Natural compounds have been proved to be useful in lowering serum cholesterol to slow down the progression of cardiovascular diseases. Pueraria lobata is employed clinically to treat cardiovascular diseases in China. In the present study, the atheroscleroprotective potential of the herb's major active compound, puerarin, was investigated by monitoring serum lipid profile and major enzyme expressions on cholesterol homeostasis in Sprague-Dawley rats fed with control diet, hypercholesterolmic diet or hypercholesterolmic diet plus administration of puerarin (300 mg/kg/day, p.o.) for 4 weeks. Puerarin markedly attenuated the increased total cholesterol induced by hypercholesterolmic diet in both serum and liver. It caused a significant reduction in the atherogenic index. Expression of mRNA for hepatic 7alpha-hydroxylase (CYP7A1) was significantly enhanced but not for those of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and lanosterol 14alpha-demethylase (CYP51). To further explore the atheroscleroprotective potential of puerarin, acetylcholine induced endothelium-dependent vasorelaxation and endothelial nitric oxide synthase (eNOS) expression on isolated thoracic aortas were analyzed. Animals administered with puerarin suppressed the hypercholesterolemic diet induced impairment of eNOS expression, whereas there was no significant difference in the endothelium-dependent vasorelaxation among various groups of animals. These data indicated that puerarin reduced the atherogenic properties of dietary cholesterol in rats. Its hypocholesterolemic function may be due to the promotion of cholesterol and bile acids excretion in liver. Whether puerarin targets directly on cholesterol homeostasis or both cholesterol homeostasis and endothelial function remains to be determined.

Publication Types: PMID: 16472823 [PubMed - indexed for MEDLINE]

112. [Effects of sucrose and light on the growth and production of secondary metabolites in Pueraria phaseoloides hairy roots]

[Article in Chinese]

He HJ, Liang P, Shi HP.

Guangdong Key Lab of Biotechnology for Plant Development, College of Life Science, South China Normal University, Guangzhou 510631, China.

Effects of sucrose concentrations and light on the growth and production of total isoflavones and puerarin in Pueraria phaseoloides hairy roots cultured onto solid MS media supplemented with 1%, 3%, 5%, 7% and 9% sucrose, respectively, were investigated. The results showed that among the sucrose concentrations tested, 3% sucrose in the medium enhanced the growth and stimulated accumulation of total isoflavones and puerarin in P. phaseoloides hairy roots, After cultured for 20 days, the biomass of hairy roots reached 0.48 g (DW dry weight)/flask and its contents of total isoflavones and puerarin were 25.44 mg/g (DW) and 11.64 mg/g (DW), respectively. In comparison with 3% sucrose, the dry weight proliferation of hairy roots cultured with 5% sucrose was increased by 7.0%, while cultured with 1%, 7% and 9% sucrose, the dry weight proliferation of hairy roots was decreased by 62.4%, 42.8% and 65.3%, their total isoflavones content was decreased by 57.4%, 13% and 33.4% and their puerarin content was decreased by 47.9%, 15.8% and 35.1%; but their content of total soluble sugars was increased 0.52, 1.45 and 1.54 times, respectively. Compared with hairy roots in blue light and white light, the biomass of hairy roots cultured in the dark for 30 days was 0.83 g (DW)/flask and was increased by 37.1% and 23.3%, respectively. The content of total isoflavones in hairy roots cultured in white light was as much as 1.15 times and 1.19 times that in blue light and in the dark, respectively. It was also observed that hairy roots cultured in blue light and white light partly became light green and that blue light could inhibit accumulation of puerarin in hairy roots and the puerarin content in hairy root cultured in white light and in the dark were 1.61 times and 1.52 times that in blue light, respectively.

Publication Types: PMID: 16468362 [PubMed - in process]

113. Puerariae radix promotes differentiation and mineralization in human osteoblast-like SaOS-2 cells.

Huh JE, Yang HR, Park DS, Choi DY, Baek YH, Cho EM, Cho YJ, Kang-Il K, Kim DY, Lee JD.


Oriental Medicine Research Center for Bone & Joint Disease, Kyung Hee University, 1 Hoegidong, Dongdaemungu, Seoul 130-701, Republic of Korea.

Puerariae radix (PR) is a traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify whether Puerariae radix extract induces osteogenic activity in human osteoblast-like SaOS-2 cells. Puerariae radix had no effect on the viability of osteoblastic cells, and dose-dependently increased alkaline phosphatase (ALP) activity. Puerariae radix markedly increased mRNA expression for vascular endothelial growth factor (VEGF), osteocalcin (OCN), osteopontin (OPN), and type I collagen (Col I) in SaOS-2 cells. Extracellular accumulation of proteins such as VEGF and Col I was increased in a dose-dependent manner. Also, Puerariae radix significantly induced mineralization in the culture of SaOS-2 cells. In conclusion, this study showed that Puerariae radix had no effect on viability, but enhanced ALP activity, VEGF, bone matrix proteins such as OCN, OPN, and Col I, and mineralization in SaOS-2 cells. These results propose that Puerariae radix can play an important role in osteoblastic bone formation, and may possibly lead to the development of bone-forming drugs.

Publication Types: PMID: 16455216 [PubMed - indexed for MEDLINE]

114. Mutagenicity of Pueraria mirifica Airy Shaw & Suvatabandhu and antimutagenicity of Thunbergia laurifolia Linn.

Saenphet K, Kantaoop P, Saenphet S, Aritajat S.

Biology Department, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand. K_Saenphet@yahoo.com

Thunbergia laurifolia Linn has been reputed to have antitoxic effects for all toxic substances. In this present study, we evaluated its effect against the mutagenicity induced by aqueous extracts from Pueraria mirifica Airy Shaw & Suvatabundhu in male rats. The formation of micronuclei in polychromatic erythrocytes was induced by oral administration of an aqueous extract of P. mirifica at the doses of 400, 600, and 800 mg/kg to the rats for 30 days. The results were that the extracts of P. mirifica at doses of 600 and 800 mg/kg acted as a mutagenic agent by inducing higher frequencies of micronuclei as compared to the controls. For the antimutagenic test, P. mirifica extract at a dose of 600 mg/kg (minimal effective dose) was mixed with fresh and dried extracts of T. laurifolia in proportions of 7:3 and 1:1, respectively. The results of 4-week-treatment indicated that aqueous extracts of T. laurifolia, prepared by both fresh and dry methods, could significantly inhibit the induction of micronuclei as induced by P. mirifica. It could be concluded from the results that, under certain circumstances, T. laurifolia exhibits a significant antimutagenic activity. The use of P. mirifica and T. laurifolia as fusion herbal medicines is suggested.

PMID: 16438216 [PubMed - indexed for MEDLINE]

115. Three indigenous Thai medicinal plants for control of Aedes aegypti and Culex quinquefasciatus.

Lapcharoen P, Apiwathnasorn C, Komalamisra N, Dekumyoy P, Palakul K, Rongsriyam Y.

Insecticide Research Unit, Department of Medical Entomology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. neparichart@yahoo.com

The potential larvicidal activity and insect growth regulator (IGR) properties of three selected indigenous medicinal Thai plants were tested against two species of mosquito with special reference to the late 3rd and early 4th instar larvae (L3 and L4, respectively). In case of larvicidal activity, Thevetia peruviana was the most potent, followed by Pueraria mirifica, and Butea superba was the least effective. In all cases, the late 3rd instar was more susceptible than the early 4th instar larvae, and the 48-hours exposure yielded more potent larvicidal activity than 24-hours exposure. However, at sublethal dosages, both P. mirifica and B. superba showed some dispersed effects interfering with ecdysis. A variety of toxic effects were observed and recorded in eight categories according to the stage of metamorphosis when death occurred. P. mirifica rendered the main deleterious effects in the pupa-adult period in both instar of Ae. aegypti and Cx. quinquefasciatus, whereas B. superba showed highest effect in black-pupa period of the late 3rd instar larval stage. The results were reversed for the early 4th instar larvae of both species of mosquito as the main effect appeared in the pupa-adult category. The overall results indicated that T. peruviana did not show any IGR properties; whereas, P. mirifica and B. superba seemed to exhibit the juvenile hormone type activity which resulted in abnormal death at various stages of development. B. superba was more promising than P. mirifica, and Ae. aegypti was about 2 times more susceptible than Cx. quinquefasciatus. In addition, L3 was always more susceptible than L4 with both mosquito species.

Publication Types: PMID: 16438204 [PubMed - indexed for MEDLINE]

116. Daidzein activates choline acetyltransferase from MC-IXC cells and improves drug-induced amnesia.

Heo HJ, Suh YM, Kim MJ, Choi SJ, Mun NS, Kim HK, Kim E, Kim CJ, Cho HY, Kim YJ, Shin DH.

Jeonnam Innovation Agency, Jeonnam, Korea.

The choline acetyltransferase (ChAT) activator, which enhances cholinergic transmission via an augmentation of the enzymatic production of acetylcholine (ACh), is an important factor in the treatment of Alzheimer's disease (AD). Methanolic extracts from Pueraria thunbergiana exhibited an activation effect (46%) on ChAT in vitro. Via the sequential isolation of Pueraria thunbergiana, the active component was ultimately identified as daidzein (4',7-dihydroxy-isoflavone). In order to investigate the effects of daidzein from Pueraria thunbergiana on scopolamine-induced impairments of learning and memory, we conducted a series of in vivo tests. Administration of daidzein (4.5 mg/kg body weight) to mice was shown significantly to reverse scopolamine-induced amnesia, according to the results of a Y-maze test. Injections of scopolamine into mice resulted in impaired performance on Y-maze tests (a 37% decreases in alternation behavior). By way of contrast, mice treated with daidzein prior to the scopolamine injections were noticeably protected from this performance impairment (an approximately 12%-21% decrease in alternation behavior). These results indicate that daidzein might play a role in acetylcholine biosynthesis as a ChAT activator, and that it also ameliorates scopolamine-induced amnesia.

Publication Types: PMID: 16428827 [PubMed - indexed for MEDLINE]

117. Reversal of chemical-induced liver fibrosis in Wistar rats by puerarin.

Zhang S, Ji G, Liu J.

School of Pharmacy, East China University of Science and Technology, Mailbox 268, Shanghai 200237, China.

Puerarin is a major isoflavonoid compound isolated from Pueraria lobata, an edible vine used widely for various medicinal purposes. It has been used for centuries in China to counteract alcohol intoxication. However, the effects of puerarin on chemical-induced liver fibrosis have not been reported. In the present study, we investigated the effects of puerarin on liver fibrosis in Wistar rats induced by alcohol plus carbon tetrachloride administration. Liver fibrosis was produced in rats by treatment with a mixture (50% alcohol, 8 g/kg per day; corn oil, 2 g/kg per day; pyrazole, 24 mg/kg per day; ig) once a day and by intraperitoneal injection of 0.25 ml/kg of a 25% solution of carbon tetrachloride in olive oil twice a week for 8 weeks. After 8 weeks, treatment with puerarin (0.4 and 0.8 g/kg ig, daily for 4 weeks) was conducted to examine its therapeutic effects. At the same time, the model group and treatment group continued to receive the chemical mixture, while the control group received saline instead of the chemical mixture. Upon pathological examination, the puerarin-treated rats significantly reversed the symptoms of liver fibrosis and other hepatic lesions. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as indexes of hepatic cell disruption, were reduced with puerarin treatment, whereas no significant effect was discovered in the levels of alkaline phosphatase (ALP) and gamma-glutamyltransferase (GGT) activities. A significant increase in apoptosis of activated hepatic stellate cell (HSC) was found by flow cytometric analysis of the hepatic tissues. And the expression of bcl-2 mRNA was down-regulated after puerarin administration. Consequently, all these results showed that puerarin could effectively reverse chemical-induced liver fibrosis in experimental rats, via the recovery of hepatic injury as well as the induction of apoptosis in activated HSC.

PMID: 16426832 [PubMed - indexed for MEDLINE]

118. Determination of isoflavones in dietary supplements containing soy, Red Clover and kudzu: extraction followed by basic or acid hydrolysis.

Delmonte P, Perry J, Rader JI.

Office of Nutritional Products, Labeling and Dietary Supplements, Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, MD 20740, USA. pierluigi.delmote@cfsan.fda.gov

Isoflavones are phytochemicals found in many plants. Because of their structural similarity to beta-estradiol, health benefits of isoflavones have been evaluated in age-related and hormone-dependent diseases. Dietary supplement preparations contain extracts from soy, Red Clover and kudzu. Soy products contain primarily genistein, daidzein, and glycitein, while Red Clover products contain primarily formononetin and biochanin A. Kudzu extracts contain puerarin and daidzein among other components. Previous methods of analysis focused on the determination of isoflavones from a single botanical source, while dietary supplements are often a blend of extracts from different plants. We developed a method for the analysis of isoflavones in dietary supplements regardless of their botanical composition, using HPLC-PDA because of its applicability to routine analysis. Isoflavones are found as free compounds, glucoside derivatives, 6''-O-malonyl-beta-d-glucoside and 6''-O-acetyl-beta-d-glucoside derivatives. In this study, the samples were extracted at room temperature with 50:50 (v/v) MeCN/water, and then analyzed before and after hydrolyzing the isoflavones by acid or basic digestion. 2'-Methoxy-flavone and 6-methoxy-flavone were used as internal standards and were added together to every sample. Daidzein, glycitein, genistein, puerarin, calycosin, pratensein, pseudobaptigenin, formononetin, biochanin A and prunetin were among the isoflavones determined.

PMID: 16413562 [PubMed - indexed for MEDLINE]

119. Traditional Chinese Medicine improves dysfunction of peroxisome proliferator-activated receptor alpha and microsomal triglyceride transfer protein on abnormalities in lipid metabolism in ethanol-fed rats.

Kwon HJ, Hyun SH, Choung SY.

Department of Hygienic Chemistry, College of Pharmacy, Kyung Hee University, Seoul 130-701, Korea.

We report the effects of Traditional Chinese Medicine (TCM) on alcohol-induced fatty liver in rats. TCM consists of Astragalus membranaceus, Morus alba, Crataegus pinnatifida, Alisma oriental, Salvia miltiorrhiza and Pueraria lobata. The rats were separated randomly into five groups; the CD group (n=10), which was fed a control diet for 10 weeks, the ED group (n=10), which was fed an isocaloric liquid diet containing ethanol for 10 weeks and given daily oral doses of TCM (0.222 g/kg/day; TCM222, 0.667 g/kg/day; TCM667, and 2.000 g/kg/day; TCM2000, n=10, respectively) over the last four weeks of the study. The ED group developed fatty livers, as determined by their lipid profiles and liver histological findings. Compared with the control group, liver/body weight, plasma triglyceride (TG) and total cholesterol (TC), liver TG and TC, plasma alanine aminotransferase (ALT) and aspartic aminotransferase (AST) significantly increased in the ED group. Also, free fatty acids (FFA) levels increased in both plasma and liver during the administration of ethanol. On the other hand, when rats were administrated with TCM, their liver/body weight, plasma TG, TC and FFA, liver TG, TC and FFA, plasma ALT and AST decreased significantly and the degree of hepatic lipid droplets was markedly improved compared with those in the ED group. Proper function of the peroxisome proliferator-activated receptor alpha (PPARalpha) is essential for the regulation of hepatic fatty acid metabolism. Microsomal triglyceride transfer protein (MTP) is essential for the secretion of triglycerides from the liver. mRNAs for PPARalpha and MTP were reduced in the livers of ethanol-fed rats. TCM restored the mRNA levels of PPARalpha and MTP, and prevented development of fatty livers in ethanol-fed rats. Impairment of PPARalpha and MTP function during ethanol consumption contributes to the development of alcohol-induced fatty liver, which can be overcome by TCM.

PMID: 16410638 [PubMed - indexed for MEDLINE]

120. [Determination of copper, zinc, iron, calcium and magnesium in pueraria lobata ohwi by FAAS]

[Article in Chinese]

Han P, Liu LE, Liu J, Zhang HQ, Wu YM, Wu YJ, Yu F, Wu DS.

College of Public Health, Zhengzhou University, Zhengzhou 450052, USA.

The samples of pueraria lobata ohwi were digested by HNO3 + HClO4. Copper, zinc, iron, calcium and magnesium in pueraria lobata ohwi from Da Bieshan were successfully determined by flame atomic absorption spectrophotometry (FAAS). The results showed that there are rich grand elements, such as Ca and Mg, and profitable elements, such as trace elements Cu, Zn and Fe, in pueraria lobata ohwi. The content sequence of metal elements is as follows: Copper: leaves > flowers > rattans > roots; Zinc: leaves > flowers > rattans > roots; Iron: roots > rattans > leaves > flowers; Calcium: leaves > roots > flowers > rattans; Magnesium: flowers > roots > leaves > rattans. The results provided useful data for discussing the relationship between the metal elements in pueraria lobata and hypertension diseases. ohwi and the cure for cerebrovascular diseases.

Publication Types: PMID: 16379303 [PubMed - in process]

121. Natural medicines for alcoholism treatment: a review.

Xu BJ, Zheng YN, Sung CK.

Department of Food Science and Technology, College of Agriculture and Biotechnology, Chungnam National University, Daejon, South Korea.

Alcoholism is a serious problem throughout the world. The development of alcoholism remedies have medical, social and economical significance. In view of the pitfalls of psychological dependence and adverse behavioural effects of synthetic drugs, the development of low toxicity and high efficiency medicines derived from natural products exhibits expansive market prospects. Based on these considerations, we summarize briefly folk application of traditional hangover remedies and clinical application of herbal complex and patent medicines for alcoholism treatment. We have reviewed the effects of natural medicines on intake, absorption and metabolism of alcohol, as well as the protective effects on alcohol-induced acute and chronic tissue injury.

Publication Types: PMID: 16361209 [PubMed - indexed for MEDLINE]

122. [Separation and identification of isoflavonoids in Pueraria lobata extracts and its preparations by reversed-phase capillary liquid chromatography coupled with electrospray ionization quadrupole time of flight mass spectrometry]

[Article in Chinese]

Tian H, Wang H, Guan Y.

Dalian Institute of Chemical Physics, The Chinese Academy of Sciences, Dalian 116023, China.

Isoflavonoids in Pueraria lobata extract and its preparations were separated and identified by reversed-phase capillary liquid chromatography (RP-CapLC) coupled with photodiode array (PDA) detector and negative electrospray ionization quadrupole time of flight (Q-TOF) mass spectrometry. The separation was performed on a 150 mm x 0. 32 mm i. d. , 5 microm C18 capillary column, using mobile phase of 0.1% aqueous trifluoroacetic acid solution and acetonitrile containing 0.1% trifluoroacetic acid under gradient elution. The product ion spectra of the deprotonated ions allowed for the identification of puerarin, daidzin and daidzein in the sample. Puerarin was found to be the most abundant component in the extract (about 13%, mass fraction) and its preparations (19.28 - 24.34 mg per tablet). The structures of trace amount of unknown isoflavonoids were deduced based on the spectra of known compounds. They were proposed to be 3'-methoxypuerarin and 3'-methoxydaidzin.

Publication Types: PMID: 16350789 [PubMed - in process]

123. Altered oral absorption of alcohol by combined aqueous extracts of four herbal plants in rats.

Shin BS, Jun H, Lee DE, Lee KR, Park ES, Yoo SD.

College of Pharmacy, Sungkyunkwan University, Changan-gu, Suwon, Kyonggi-do, Korea.

This study examined the effect of combined aqueous extracts (BHR) of Ginko biloba, Mentha arvensis var. piperascens, Citrus unshiu, and Pueraria lobata var. chinensis on oral absorption of alcohol in rats. The rats were pretreated with BHR, placebo solution identical to BHR without the herbal extract, and isotonic saline. Alcohol was administered orally at 1- and 3-g/kg doses and the absorption profiles were compared. After oral administration of 1-g/kg doses, mean area under the curve (AUC) and C(max) values were significantly reduced in BHR-treated rats (16.1 +/- 10.0 and 0.3 +/- 0.1 mg/ml, respectively) as compared with saline-treated (37.9 +/- 14.4 and 0.7 +/- 0.7 mg/ml, respectively) and placebo solution-treated (63.0 +/- 46.4 and 0.7 +/- 0.4 mg/ml, respectively) rats. Similarly, after administration of 3-g/kg doses, mean AUC and C(max) values in BHR-treated rats (188.1 +/- 119.7 mg(.)min/ml and 1.0 +/- 0.4 mg/ml) were significantly reduced over those in saline-treated rats (571.4 +/- 512.4 mg(.)min/ml and 1.8 +/- 0.9 mg/ml, respectively). The relative oral bioavailability of alcohol calculated as the ratio of AUC(BHR)/AUC(Saline) was 42.5% and 32.9% at 1- and 3-g/kg doses, respectively. The reduced serum alcohol levels as well as the reduced AUC and C(max) after pretreatment with BHR appear to be a result of a reduced systemic absorption not due to an increased metabolic clearance.

Publication Types: PMID: 16326435 [PubMed - indexed for MEDLINE]

123. [Effect of pueraria crude extreact and puerarin on ethanol-induced expression of heat shock protein 70 in embryonic mouse hippocampal cultures]

[Article in Chinese]

Han P, Wu DS, Li WJ, Yu ZL, Wang Q.

Huaxi Public Health School, Sichuan University, Chengdu 610041, China.

OBJECTIVE: To study if the Pueraria crude extreact (CP) and standard preparation of pure puerarin (SP) possess the same neuroprotective effects on the expression of heat shock protein (HSP) 70 in the embryonic mouse hippocampal cells. METHODS: The hippocampus of 18-days-old mouse embryo was taken out and suspension of single cells was cultured. Ethanol was added to cause HSP70 mRNA expression. Solvent, ethanol of different concentrations (50, 200, and 300 mmol/L), SP + ethanol, and SP + ethanol were added respectively. Western blotting was used to detect the expression of the expression of HSP70 mRNA. RESULTS: Ethanol of different concentrations increased the expression of HSP70 mRNA and the protein in comparison with the solvent control group. SP and CP inhibited the expression of HSP70 mRNA and protein. CONCLUSION: With identical effect of anti-oxidative stress, both SP and CP inhibit the increase of expression of HSP70 mRNA and protein, thus demonstrating I vitro anti-oxidative neuroprotection.

Publication Types: PMID: 16324368 [PubMed - indexed for MEDLINE]

125. Puerarin protects rat pancreatic islets from damage by hydrogen peroxide.

Xiong FL, Sun XH, Gan L, Yang XL, Xu HB.

College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.

The protective effect of puerarin, an isoflavone purified from Chinese herb radix of Pueraria lobata, on hydrogen peroxide (H(2)O(2))-induced rat pancreatic islets damage was investigated. Exposure of islets to 500 microM H(2)O(2) could cause a significant viability loss and an increase in apoptotic rate. Pretreatment of islets with puerarin for 48 h resulted in a reduction in viability loss and apoptotic rate. 100 microM puerarin significantly inhibited the apoptosis of islets induced by H(2)O(2). In addition, preincubation with puerarin could restore the H(2)O(2)-induced decrease in basal and glucose-stimulated insulin secretion in pancreatic islets. Puerarin was also found to inhibit the free radicals production induced by H(2)O(2) and to increase catalase and superoxide dismutase (SOD) activities in the isolated pancreatic islets. These results suggest that puerarin can protect islets against oxidative stress probably due to stimulating the activities of the antioxidant enzymes. Puerarin may be effective in preventing islet cells from the toxic action of reactive oxygen species in diabetes.

Publication Types: PMID: 16321378 [PubMed - indexed for MEDLINE]

127. [Potential antiviral drug Pueraria crude extract and puerarin protect against ethanol-induced cytotoxicity in embryonic mouse hippocampal cultures]

[Article in Chinese]

Han P, Li J, Li WJ, Yu ZL, Wang Q, Wu DS.

Huaxi School of Public Health, Sichuan University, Chengdu 610041, China.

OBJECTIVE: To examine whether Chinese medical herb Pueraria crude extract (CP) and standard of pure puerarin (SP) possess the same neuroprotective effects during concomitant ethanol (EtOH) treatment. METHODS: Hippocampus cultures were prepared from mice at gestational age of 18 day. Cell viability was measured by MTT assay. RT-PCR was employed to determine mRNA expression of superoxide dismutase (SOD). RESULTS: As measured by MTT assay, supplementation with 15 mg/L CP or 10 mg/L SP afforded neuroprotection against all EtOH concentrations (50, 200 and 350 mmol/L, respectively) in embryonic hippocampal culture system. In addition, both 15 mg/L CP and 10 mg/L SP could decrease expression of SOD at mRNA level. CONCLUSION: This study suggests that CP and SP could decrease oxidative stress induced by ethanol treatment by the decreased expression of SOD at mRNA level, and demonstrates antioxidative neuroprotective effect of CP and SP against developmental ethanol exposure in vitro.

Publication Types: PMID: 16261207 [PubMed - indexed for MEDLINE]

128. Suppression of arachidonic acid metabolism and nitric oxide formation by kudzu isoflavones in murine macrophages.

Jun M, Hong J, Jeong WS, Ho CT.

Department of Food Science, Rutgers University, 65 Dudley Road, New Brunswick, NJ 08901-8520, USA.

Inhibitory effect of kudzu isoflavones on arachidonic acid metabolism and nitric oxide (NO) production in lipopolysaccharide activated RAW 264.7 macrophages were investigated. Isoflavone aglycones, such as daidzein, genistein, biochanin A, and formononetin significantly suppressed arachidonic acid release (50 microM). Biochanin A, which displayed the most active inhibition on arachidonic acid release in HT-29 human colon cancer cells, exhibited its most potent suppression in RAW 264.7 cell (by 86%) without showing cytotoxicity. However, isoflavone glucosides, puerarin and daidzin, showed lower inhibitory activities on the release of arachidonic acid and its metabolites. In NO formation, biochanin A showed marked inhibition, by 62% (50 microM), followed by genistein, daidzein, formononetin, and daidzin, 56, 39, 33, and 8%, respectively. 5,7-Dihydroxyl group in the A-ring of isoflavones could be a key functional group responsible for the strong inhibitory activity of biochanin A and genistein on NO production. These activities may contribute to the antiinflammatory and anticarcinogenic properties of kudzu isoflavones.

PMID: 16254887 [PubMed - indexed for MEDLINE]

129. Puerariae radix prevents bone loss in castrated male mice.

Wang X, Wu J, Chiba H, Yamada K, Ishimi Y.

Division of Applied Food Research, National Institute of Health and Nutrition, Tokyo 162-8636, Japan.

Puerariae radix (PR) is one of the earliest and most important crude herbs used in Chinese medicine for various medicinal purposes. PR contains a high amount of isoflavonoids, such as daidzein and genistein, which are known to prevent bone loss induced by estrogen deficiency. We have demonstrated that PR not only completely prevents bone loss but also significantly increases the bone mass at high doses in ovariectomized mice without exhibiting estrogenic action in the uterus. In this study, we examined whether PR exhibits effects on bone loss in androgen-deficient male mice similar to estrogen-deficient female mice. Male mice were orchidectomized (ORX) and fed a diet containing low, middle, and high doses (5%, 10%, and 20% of diet, respectively) of PR or normal diet with subcutaneous administration of 17beta-estradiol (E(2), 0.03 microg/d; Sigma, St Louis, Mo), for 4 weeks. In ORX mice, the seminal vesicle weight decreased markedly, and it was not affected by the administration of any doses of PR and E(2). The bone mineral density (BMD) of the whole femur was significantly decreased by ORX, and the decrease in BMD was completely prevented by intake of the diet with the low dose of PR. Intake of the diet with the middle dose of PR further normalized BMD in ORX mice. Furthermore, the high dose of PR administration (PR20) significantly increased BMD in ORX mice, and the potency was similar to that of E(2). Morphometric analysis of the femoral metaphysis showed that intake of the diet with the low dose of PR completely prevented the decrease in bone volume/tissue volume and trabecular number and restored the increase in trabecular separation in ORX mice. In addition, intake of the diet with the high dose of PR further increased bone volume/tissue volume and trabecular number and decreased trabecular separation in ORX mice. These results propose the possibility that estrogenic Chinese herbs such as PR can be one of the candidates for the treatment or prevention of osteoporosis in elderly men with hypogonadism.

Publication Types: PMID: 16253645 [PubMed - indexed for MEDLINE]

130. High-performance liquid chromatography-tandem mass spectrometry for identification of isoflavones and description of the biotransformation of kudzu root.

Zhang Y, Xu Q, Zhang X, Chen J, Liang X, Kettrup A.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, PR China.

High-performance liquid chromatography-tandem mass spectrometry has been used to identify isoflavone aglycones and glycosides in kudzu root. Fourteen isoflavones were detected. Among these, six were identified by comparison with authentic standards. Tentative identifications of the other isoflavones are based on UV spectra, mass spectra of protonated and deprotonated molecules, and MS-MS data. Several are reported for the first time in kudzu root. The bioactivity and bioavailability of isoflavone aglycones are usually greater than those of their glycosides. To improve the bioavailability of kudzu root isoflavones, crude beta-glycosidases prepared from microbes were used to hydrolyze the isoflavone glycosides. Several MS modes are combined not only to identify the isoflavones in kudzu root, but also to describe the biotransformation of kudzu root isoflavone glycosides. It is also proved that crude beta-glycosidases have high selectivity toward the O-glycosides of isoflavones.

Publication Types: PMID: 16237543 [PubMed - indexed for MEDLINE]

131. Determination of 12 pueraria components by high-performance liquid chromatography-mass spectrometry.

Lin CC, Wu CI, Sheu SJ.

Department of Chemistry, National Taiwan Normal University, Taipei, Taiwan, P.R. China. chingchekimo@yahoo.com.tw

Puerariae radix, a commonly used Chinese herb drug derived from the dried root of legume plant, contains a series of isoflavones as its chief pharmacologically active constituents. Using 12 pueraria components as markers, an LC-UV-MS method requiring less than 60 min, was developed for estimating the quality of pueraria samples within 60 min. Extracts were analyzed using a Cosmosil 5C18-MS column, by gradient elution with an aqueous solution of acetic acid and methanol-ACN at a flow-rate of 1.0 mL/min. Peaks were detected at 254 nm and each peak was identified by LC/MS. The reproducibilities (RSD) of this proposed method, on the basis of peak-area ratios from six replicate injections, were 0.93-1.42% (intraday) and 1.05-1.63% (interday) at a detection limit of 0.12-0.78 microg/mL. Most of the markers used in this study can be classified, respectively, into three major categories, namely, isoflavones, O-glycosidic isoflavones, and C-glycosidic isoflavones. The structures of the compounds were determined from LC-MS fragmentation data and data from the literature.

Publication Types: PMID: 16224974 [PubMed - indexed for MEDLINE]

132. Amelioration effects of traditional Chinese medicine on alcohol-induced fatty liver.

Kwon HJ, Kim YY, Choung SY.

Department of Hygienic Chemistry, College of Pharmacy, Kyung Hee University, Seoul 130-701, South Korea.

AIM: To examine the effects of traditional Chinese medicine (TCM) on alcohol-induced fatty liver in rats. TCM consists of Astragalus membranaceus, Morus alba, Crataegus pinnatifida, Alisma orientale, Salvia miltiorrhiza, and Pueraria lobata. METHODS: The rats were separated randomly into five groups. One (the CD group) was fed a control diet for 10 wk, another (the ED group) fed an ethanol-containing isocaloric liquid diet for 10 wk, and the last three (the TCM group) were fed an ethanol-containing isocaloric liquid diet for 10 wk and dosed orally with TCM (222 mg/kg.d, TCM222; 667 mg/kg.d, TCM667; and 2 000 mg/kg.d, TCM2000, respectively) weekly during the last 4 wk. RESULTS: ED group developed fatty liver according to lipid profile and liver histological findings. Compared with the control group, liver/body weight, serum triglyceride (TG) and total cholesterol (TC), liver TG and TC, serum alanine aminotransferase (ALT) and aspartic aminotran-sferase (AST) significantly increased in the ED group. Whereas, in the rats administered with TCM, liver/body weight, serum TG and TC, liver TG and TC, serum ALT and AST were significantly decreased, and the degree of hepatic lipid droplets was markedly improved compared with those in the ED group. CONCLUSION: TCM treatment causes significant reduction in alcohol-induced lipid hepatic accumulation, reversing fatty liver and liver damage, and can be used as a remedy for alcoholic fatty liver.

PMID: 16222745 [PubMed - indexed for MEDLINE]

134. Evaluation of Nigerian animal feeds by particle-induced X-ray emission.

Olabanji SO, Olubunmi P, Ceccato D, Buoso MC, De Poli M, Moschini G.

Istituto Nazionale di Fisica Nucleare, Laboratori Nazionali di Legnaro, Padova, Italy.

There is need to evaluate the locally available animal feeds in Nigeria so as to be able to combine them in acceptable proportions to the animals to achieve the desired growth rate. The technique of particle-induced X-ray emission (PIXE) was employed for the evaluation of these locally available animal feeds, which include Panicum maximum (Guinea grass), Cynodon plectostachyum (grass), Leucaena leucephala (legume), Calopogonium mucunoides (legume), Gliricidia sepium (legume), Euphorbia polychrome (legume), Pueraria phaseloides (legume), and Centrosema pubescens (legume). The proton beam delivered by the 2.5-MV AN 2,000 Van de Graaff accelerator at the International Centre for Theoretical Physics Laboratori Nazionali di Legnaro (LNL), Padova, Italy was used for the PIXE measurements. Twenty-one different elements were detected at various concentrations and their nutritional effects on different animals are discussed.

Publication Types: PMID: 16217142 [PubMed - indexed for MEDLINE]

135. Effects of an aqueous extract of Puerariae flos (Thomsonide) on impairment of passive avoidance behavior in mice.

Yamazaki T, Yaguchi M, Nakajima Y, Hosono T, Niiho Y, Hibi Y, Kinjo J, Nohara T.

Tsukuba Research Institute, Ohat's Isan Co. Ltd., Ushiku, Ibaraki 300-1231, Japan. t_yamazaki@ohta-isan.co.jp

The effects of an aqueous extract of Puerariae flos (Thomsonide) on ethanol-induced learning and memory impairment and scopolamine-induced amnesia were investigated. Thomsonide exerted an ameliorating effect on the impairment of both memory registration and memory retrieval induced by ethanol. These results indicate that Thomsonide has an antiamnesic effect on the central nervous system in alcoholic intoxication and support the traditional use of Puerariae flos for the treatment of alcoholic intoxication. Thomsonide also improved the scopolamine-induced impairment of memory registration in passive avoidance behavior in mice. The results of this study suggest that it may be possible to use Thomsonide for the treatment of age-related memory impairment and dementia.

PMID: 16125021 [PubMed - indexed for MEDLINE]

136. Protection by puerarin against MPP+-induced neurotoxicity in PC12 cells mediated by inhibiting mitochondrial dysfunction and caspase-3-like activation.

Bo J, Ming BY, Gang LZ, Lei C, Jia AL.

School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian 116024, China. bojiang0411@yahoo.com.cn

Puerarin, a main isoflavone glycoside distributed in Pueraria lobata (Willd.) Ohwi, showed inhibitory activity on H2O2-induced PC12 cells damage in our previous work. However, there is insufficient evidence in protective mechanism of puerarin, especially that relating to the mitochondrial function. In this study, when cells were pretreated with puerarin prior to 0.4 mM MPP+, protective roles were accompanied by a reduction of cell viability loss, morphological changes of apoptosis and apoptotic rate. To explore the protective mechanism of puerarin in MPP+-induced PC12 cells, mitochondrial function and caspase-3-like activity were measured. The results indicated that puerarin inhibited the release of mitochondrial cytochrome c to cytosol and the loss of mitochondrial membrane potentials. In addition, puerarin also reduced MPP+-induced caspase-3-like activation. Taken together, the above results suggest that pretreatment of PC12 cells with puerarin could block MPP+-mediated apoptosis by mitochondria-dependent caspase cascade.

PMID: 16112764 [PubMed - indexed for MEDLINE]

137. [Protective effect of purariae isoflavone on apoptosis cells of nasal mocosas in ovariectomized rats]

[Article in Chinese]

Qi BM, Cai CC, Deng XM, Zhang L.

The First People's Hospital of Yunnan Province, Kunming 650001, China. qibenming@21cn.com

[OBJECTIVE: To study the protective effect of purariae isoflavone on apoptosis cells of atrophic nasal mucosas in ovariectomized rats. METHOD: 60 rats were divided into four groups as control, ovariectomized, ovariectomized + nylestriol (O + N) and ovariectomized + purariae isoflavone (O + P), each with 15 rats. Earlier apotosis cells of mucosas taken from nasal septum were measured with flow cytometry. RESULT: Compared with control group, and the number of apoptosis cells of mucosas increased after being ovariectomized,and the number of apoptosis cells of mucosas in O + N and O + D group didn't change. CONCLUSION: Nylestriol and purariae isoflavone might have effects on protecting cells of mucosas from lacking of estrogen by decreasing apoptosis cells in ovariectomized rats.

Publication Types: PMID: 16110871 [PubMed - indexed for MEDLINE]

138. Self-microemulsifying drug delivery systems (SMEDDS) for improving in vitro dissolution and oral absorption of Pueraria lobata isoflavone.

Cui S, Zhao C, Chen D, He Z.

School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, PR China.

The aim of our investigation was to develop and characterize self-microemulsifying drug delivery systems (SMEDDS) of Pueraria lobata isoflavone to improve its in vitro dissolution and oral absorption in beagle dogs. SMEDDS consisted of oil (ethyl oleate), a surfactant (Tween 80), and a cosurfactant (Transcutol P). In all the SMEDDS, the level of Pueraria lobata isoflavone was fixed at 20% w/w of the vehicle. The in vitro self-microemulsification properties and droplet size analysis of SMEDDS were studied following their addition to water under mild agitation. A pseudoternary phase diagram was constructed identifying the efficient self-microemulsification region. From these investigations, an optimized formulation was selected and its dissolution and bioavailability were compared with a tablet formulation in beagle dogs. The in vitro dissolution rate of puerarin from SMEDDS was more than threefold faster than that from Yufengningxin tablets (Pueraria lobata isoflavone tablets). A 2.5-fold increase in the relative bioavailability was observed for the SMEDDS compared with Yufengningxin tablets. The absolute bioavailability of the SMEDDS was 82.32 +/- 15.51%, which was significantly improved compared with that of Yufengningxin tablets. These results demonstrate the potential of SMEDDS as an efficient way of improving the oral absorption of Pueraria lobata isoflavone.

Publication Types: PMID: 16093200 [PubMed - indexed for MEDLINE]

139. Determination of puerarin in rat cortex by high-performance liquid chromatography after intravenous administration of Puerariae flavonoids.

Yan B, Wang W, Zhang L, Xing D, Wang D, Du L.

Laboratory of Pharmaceutical Sciences, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, People's Republic of China.

In the present study, a rapid and simple high-performance liquid chromatographic (HPLC) assay for determination of puerarin in rat cortex was developed. The analysis was carried out on a Zorbax SB-C18 column with mobile phase acetonitrile-0.5% aqueous phosphoric acid (11:89, v/v). The detection was by UV at 252 nm. The calibration curve for puerarin was linear (r=0.9999) over the concentration range 0.516-206.250 microg/mL. The limit of detection was 0.206 microg/mL (signal-to-noise ratio 3) and the limit of quantification (signal-to-noise ratio 10) was 0.516 microg/mL. Stability studies showed that puerarin was stable at temperatures of 4 degrees C in methanol for at least 30 days. The intra- and inter-day assays of puerarin from rat cortex were less than 2.5% at concentration range 0.516-206.250 microg/mL and good overall recoveries (97.4-101.7%) were found at same concentrations. The method was applied to determine the pharmacokinetic parameters and the time course of puerarin in rat cortex, following a single dosage of intravenous administration of flavonoids from Puerariae radix at 32 mg/kg of puerarin to male Wistar rats. Copyright (c) 2005 John Wiley & Sons, Ltd.

Publication Types: PMID: 16078309 [PubMed - indexed for MEDLINE]

140. Induction of apoptosis by puerarin in colon cancer HT-29 cells.

Yu Z, Li W.

School of Public Health, Zhengzhou University, Zhengzhou 450052, China. yuzengli@263.net

Puerarin was isolated from Pueraria radix and has beneficial effects on cardiovascular, neurological, and hyperglycemic disorders. The current study showed that puerarin also possessed anti-cancer properties. Methyl thiazolyl tetrazolium assay (MTT) assay revealed a dose-dependent reduction of HT-29 cellular growth in response to puerarin treatment. Apoptosis was observed following treatments ;with >or=25 microM puerarin, as reflected by the appearance of the subdiploid fraction and NDA fragmentations. We then investigated effects of puerarin on expression of apoptosis-associated genes and the results revealed an increase of bax and decreases of c-myc and bcl-2. Finally, puerarin treatment significantly increased the activation of caspase-3, a key executioner of apoptosis. These findings indicate that puerarin may act as a chemopreventive and/or chemotherapeutic agent in colon cancer cells by reducing cell viability and inducing apoptosis.

PMID: 16055262 [PubMed - indexed for MEDLINE]

141. The Neuroprotection of puerarin against cerebral ischemia is associated with the prevention of apoptosis in rats.

Xu X, Zhang S, Zhang L, Yan W, Zheng X.

Department of Biomedical Engineering, Zhejiang University, Hangzhou, P. R. China.

Previous work has shown that puerarin (Pur), extracted from the dried root of Pueraria lobata (Wild) Ohwi, increases cerebral blood flow in dogs and attenuates cerebral and spinal cord injury resulting from ischemia and reperfusion in rats and rabbits. The present study further demonstrates the neuroprotective effects of Pur on cerebral ischemic injury in rats and the mechanisms underlying the protective effects. Male Sprague-Dawley rats were subjected to middle cerebral artery occlusion (MCAo) for 50 min followed by reperfusion for 24 h. Pur (50, 100 mg/kg, i.p) was administered at the onset of MCAo. Twenty-four hours after reperfusion, neurological deficits were evaluated in Pur- and vehicle-treated rats. The infarct volume and edema ratios were assessed from stained brain slices. The results showed that Pur (100 mg/kg) markedly decreased the infarct volume by 34 % ( P < 0.01) in cerebral cortex and improved the neurological functions ( P < 0.05) after MCAo. Furthermore, flow cytometric analysis of annexin-V and PI labeling cells showed that the percentages of apoptosis and necrosis in the dorsolateral cortex were significantly reduced by 38.6 % and 28.5 % ( P < 0.01 and P < 0.05) following treatment with Pur (100 mg/kg) in MCAo rats. Caspase-3 activity, a biochemical marker of apoptosis, was significantly inhibited after treatment with Pur in the dorsolateral cortex. In agreement with this result, the expression of the X-chromosome-linked inhibitor of apoptosis protein (XIAP) was obviously up-regulated after administration of Pur (100 mg/kg), while caspase-3 gene was down-regulated in the dorsolateral cortex. These results suggest that the neuroprotection of puerarin against cerebral ischemia is associated with anti-apoptosis.

Publication Types: PMID: 16041641 [PubMed - indexed for MEDLINE]

142. The transgenic Arabidopsis plant system, pER8-GFP, as a powerful tool in searching for natural product estrogen-agonists/antagonists.

Chang FR, Hayashi K, Chua NH, Kamio S, Huang ZY, Nozaki H, Wu YC.

Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan, Republic of China. aaronfrc@kmu.edu.tw

The transgenic Arabidopsis plant system, pER8-GFP, may be used as a powerful tool in searching for natural estrogen-agonists/antagonists. Among selected plant extracts and natural products, the method was able to distinguish active extracts (e.g., Glycine max and Pueraria lobata) and pure compounds (e.g., 17beta-estradiol (1), genistein (10), and daidzein (11)) and also to distinguish effects of structural changes on activity. Thus, this rapid sensitive system was proven to be suitable for the discovery of natural products with estrogenic activity.

Publication Types: PMID: 16038533 [PubMed - indexed for MEDLINE]

143. Characterization of beta-hydroxy-beta-methylglutaryl coenzyme A reductase inhibitor from Pueraria thunbergiana.

Kim HJ, Lee DH, Hwang YY, Lee KS, Lee JS.

Department of Genetic Engineering and Bio-medicinal Resources Research Center and Department of Biology and Medicinal Science, Paichai University, Daejeon 302-735, South Korea.

This study describes the extraction and characterization of an inhibitor for beta-hydroxy-beta-methylglutaryl (HMG) coenzyme A (CoA) reductase from Pueraria thunbergiana. The maximum HMG-CoA reductase inhibitory activity (IC(50) = 79 microg) was obtained when P. thunbergiana was extracted with 70% ethanol at 30 degrees C for 12 h. After purification of the HMG-CoA reductase inhibitor by means of systematic solvent extraction, silica gel column chromatography, and HPLC, an active fraction with an IC(50) of 0.9 microg (4.25 microM) and a yield of 1.3% was obtained. The purified HMG-CoA reductase inhibitor was identified as daidzein (C(15)H(10)O(4); molecular mass, 254 Da).

Publication Types: PMID: 16028969 [PubMed - indexed for MEDLINE]

144. [Experimental study of protective effect of pueraria compound on the cerebral ischemic injury]

[Article in Chinese]

Zhao Y, Du GY, Cui HF, Cao CY, Wang XR, Zhang CY.

Institute of Chinese Materia Medica, China Academy of Traditional Chinese Medicine, Beijing 100700, China.

OBJECTIVE: To discuss the protective effects of pueraria compound on the cerebral ischemic injury. METHOD: Using the middle cerebral artery occlusion model (MCAO) in rats and cerebral ischemia-reperfusion models in gerbils and mice, we investigated the influence of pueraria compound on the brain water content and the infarct size, the cerebral apoplexy exponent, the contents of lactic acid (LA) and lipid peroxide (LPO), the activities of lactic dehydrogenase (LDH), glutathione peroxidase (GPx) and Na+ -K+ -ATPase. RESULT: Pueraria compound obviously reduced the brain water content and the infrarct size in MCAO, improved motor abilities in the cerebral ischemia-reinfusion model of gerbils, decreased the contents of LA and LPO and increased the activities of LDH, GPx and Na+ -K+ -ATPase in cerebral ischemia-reinfusion model of mice. CONCLUSION: Pueraria compound has the function of antioxidation and protective effect on ischemic brain tissue.

Publication Types: PMID: 16011106 [PubMed - indexed for MEDLINE]

145. [Effect of puerarin on the expression of Hsp70 in the rats with cerebral injury induced by acute local ischemia]

[Article in Chinese]

Pan HP, Mo XL, Yang JZ, Li LL, Huang ZL, Ye J, Huang J.

People's Hospital of Guangxi Autonomous Region, Nanning 530021, China. php1968@126.com

OBJECTIVE: To study the effect of puerarin on the expression of Hsp (heat shock protein) 70 in the rats with cerebral injury induced by acute local ischemia. METHOD: Rat model of acute local cerebral ischemia was made by ligating middle cerebral artery. The Hsp70 expression in brain tissue was detected by SP method of immunohistochemistry. RESULT: The expression of Hsp70 was significantly higher in puerarintreated rats than those in the rats with cerebral ischemia. CONCLUSION: Puerarin can enhance the level of Hsp70 expression in the rats with cerebral injury induced by acute local ischemia.

Publication Types: PMID: 16011103 [PubMed - indexed for MEDLINE]

146. Determination of puerarin in human plasma by high performance liquid chromatography.

Ma Z, Wu Q, Lee DY, Tracy M, Lukas SE.

Bio-organic and Natural Products Laboratory, McLean Hospital, Harvard Medical School, 115 Mill Street, Belmont, MA 02478, USA.

Puerarin, an isoflavone C-glycoside, has been identified as the major active component isolated from Pueraria lobata (Kudzu) responsible for suppression of alcohol drinking. In order to conduct clinical studies of Kudzu's efficacy, a method for measuring its bioavailability and pharmacokinetic profile is needed. We have developed a gradient reversed-phase HPLC system for pharmacokinetic study of puerarin in human plasma. Solid-phase extraction was performed on an abselut Nexus cartridge (60 mg/3 ml) possessing adsorbent function with a recovery of >97% and 4-hydroxybenzoic acid was used as an internal standard. The HPLC assay was performed on a YMC ODS-A column (150 mm x 4.6mm i.d., 5 microm particle size). The HPLC mobile phase consisted of methanol/0.5% acetic acid with 20-35% methanol gradient at a flow-rate of 0.8 ml/min. The UV wavelength was set at 254 nm. Calibration of the overall analytical procedure gave a linear signal (r>0.999) over a puerarin concentration range of 5-500 ng/ml in human plasma. The lower limit of quantification was ca. at 8 ng/ml of puerarin in plasma. The detection limit (defined as signal-to-noise ratio of about 3) was approximately 3 ng/ml. The preliminary pharmacokinetic study after oral administration of the Kudzu capsules containing 400mg of puerarin to a healthy volunteer confirmed that the present method was suitable for determining puerarin in human plasma.

Publication Types: .PMID: 16009605 [PubMed - indexed for MEDLINE]

147. The study of anti-metabolic syndrome effect of puerarin in vitro.

Xu ME, Xiao SZ, Sun YH, Zheng XX, Ou-Yang Y, Guan C.

Department of Biomedical Engineering, Zhejiang University (Yuquan Campus), Hangzhou 310027, PR China.

Puerarin is an isoflavone extracted from Chinese plant, Pueraria lobata (Wild.) Ohwi. It has been reported to have comprehensive pharmacological action in treatment of diabetes and cardiovascular diseases. The purpose of this study was to link the scattered effects of puerarin and to find the common mechanisms underlying. We investigated the effect of puerarin on the pivotal common pathogenic factors of metabolic syndrome, which includes obesity, Type II diabetes and cardiovascular diseases. Recently, a large body of evidence indicates that there is a complicated interplay among insulin resistance, adipocytes and endothelial dysfunction that links the abnormalities of metabolic syndrome. Results of present study showed that puerarin could potentiate insulin-induced preadipocyte differentiation, promote glucose-uptake of adipocytes that have been induced insulin resistance by high glucose, and prevent TNF-a-induced apoptosis and viability loss of endothelial cells. Furthermore, we found that these effects are probably due to promote PPARgamma expression and partly through inhibiting abnormal TNF-a-induced intracellular-free Ca(2+) accumulation of endothelial cells. Overall, our synthetical study links the comprehensive pharmacological actions of puerarin to the recognized common pathogenesis of metabolic syndrome, and provides a new insight into the mechanism of puerarin effect.

Publication Types: PMID: 16005472 [PubMed - indexed for MEDLINE]

148. Antipyretic, analgesic and muscle relaxant activities of pueraria isoflavonoids and their metabolites from Pueraria lobata Ohwi-a traditional Chinese drug.

Yasuda T, Endo M, Kon-no T, Kato T, Mitsuzuka M, Ohsawa K.

Department of Phytochemistry, Tohoku Pharmaceutical University, Sendai, Japan.

We evaluated the antipyretic, analgesic, and muscle relaxant activities of Pueraria isoflavonoids and their metabolites in mice. The glycosides daidzin and genistin significantly reduced fever induced by lipopolysaccharide (LPS). Their metabolites, daidzein and p-ethylphenol, also significantly reduced fever induced by LPS. In addition, daidzin, daidzein, dihydrodaidzein, and p-ethylphenol showed analgesic activity as assessed by the acetic acid-induced writhing test. Furthermore, equol and p-ethylphenol showed muscle relaxant activity in the rotarod and horizontal wire test. These results suggest that these compounds play a major role in the therapeutic activity of Pueraria isoflavonoids.

PMID: 15997103 [PubMed - indexed for MEDLINE]

149. Protective effect of puerariae radix on oxidative stress induced by hydrogen peroxide and streptozotocin.

Kang KA, Chae S, Koh YS, Kim JS, Lee JH, You HJ, Hyun JW.

Department of Biochemistry, College of Medicine and Applied Radiological Science Research Institute, Cheju National University, Korea.

This study evaluated the protective effect of Puerariae radix against the oxidative stress induced by hydrogen peroxide (H2O2) and streptozotocin in vitro and in vivo, respectively. The ethanol extract scavenged intracellular reactive oxygen species (ROS), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and prevented lipid peroxidation. This radical scavenging activity of the ethanol extract protected the cell viability of Chinese hamster lung fibroblast (V79-4) cells exposed to H2O2. Furthermore, this extract reduced the formation of apoptotic cells induced by H2O2, which was demonstrated by the decreased number of sub G(1) hypo-diploid cells and apoptotic cell body formation. The extract increased the activities of the cellular antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT). Administration of the extract to the streptozotocin induced diabetic rats decreased the blood glucose levels. The diabetic rats showed low activities of superoxide dismutase and catalase in the liver, and the ethanol extract increased the CAT activity. The increased level of lipid peroxidation in the diabetic rats reverted to near normal levels after being treated with the extract. This study showed that Puerariae radix was effective in the amelioration of diabetes, which may be a consequence of its antioxidant potential.

Publication Types: PMID: 15997089 [PubMed - indexed for MEDLINE]

150. [Induction of hairy roots of Pueraria phaseoloides and its culture in liquid and solid medium]

[Article in Chinese]

Shi HP, Quan H, Spiros K.

School of Life Science, South China Normal University, Guangzhou 510631, China. shihp@scnu.edu.cn

An efficient transformation system for genetic transformation of medicinal plant, Pueraria phaseoloides, which contains puerarin and daidzein with hypothermic, spasmolytic, hypotensive and anti-arrhythmic activities, by using agropine-type Agrobacterium rhizogenes ATCC 15834 was developed. Hairy roots could be obtained directly from the cut edges of petioles of leaf explants of P. phaseoloides or via callus 20 days after inoculation with agrobacterium. The percentage of rooted leaf explants 35 days after infection was about 85%. Hairy roots could have a rapid growth on solid or liquid growth regulator-free MS medium. The transformation of hairy roots was confirmed by PCR amplification of rol B and rol C genes of Ri plasmid from A. rhizogenes. To investigate the physiological difference between solid and liquid culture, the biomass (fresh weight and dry weight), the reactive oxygen species (ROS) and the total content of soluble sugar in hairy roots cultured for 15 days in solid and liquid medium were detected, respectively, by the method of fluorescence labeling of 2',7'-dichlorofluorescein diacetate (2',7'-DCFH-DA) and by the anthrone colourimetry. Compared to hairy roots in solid medium, hairy roots grew more rapidly in liquid medium but formed no callus and appeared to become brown earlier during culture. The fresh weight, the dry weight, the total content of soluble sugar and the levels of reactive oxygen species of hairy roots cultured into liquid medium MS without plant growth regulators for 15 days were 1.59 times, 1.18 times, 5.25 times and 1.16 times, respectively as much as that of hairy roots cultured onto solid medium. Our results firstly indicate that P. phaseoloides hairy roots in solid medium can utilize or metabolize more soluble sugar but produce less reactive oxygen species than that in liquid medium. This may be related to the fact that hairy roots are easier to turn brown in liquid medium than that onto solid medium. Our results have laid a foundation for defining optimum culture manner for large-scale cultivation and large-scale production of secondary metabolites of P. phaseoloides hairy roots.

Publication Types: PMID: 15969012 [PubMed - in process]

151. Protective effects of hydroxyethylpuerarin on cultured bovine cerebral microvascular endothelial cells damaged by hydrogen peroxide.

Guang HM, Zhang XM, Li YQ, Wei XB, Wang ZY, Liu HQ.

Department of Pharmacology, School of Medicine, Shandong University, Jinan 250012, China.

AIM: To observe the damages induced by hydrogen peroxide in cultured bovine cerebral microvascular endothelial cells (BCMEC) and evaluate the protective effects of hydroxyethylpuerarin on hydrogen peroxide-injured BCMEC. METHODS: BCMEC were cultured and transferred into modified Eagle medium (MEM). The viability of cells was detected by MTT assay. Cell injury was determined by lactate dehydrogenase (LDH) activity in the extracellular medium. Flow cytometry was employed to observe the occurrence of apoptosis. Morphologic changes of cells were visualized under phase contrast and electron microscopes. RESULTS: Hydrogen peroxide (200 micromol x L(-1) for 4 hours) inhibited the viability of cultured BCMEC and stimulated LDH release. Hydrogen peroxide (100 micromol x L(-1) for 4 hours) induced the occurrence of apoptosis. Hydroxyethylpuerarin was shown to increase the survival rate and decrease the activity of LDH of BCMEC damaged by hydrogen peroxide. Hydroxyethylpuerarin was also found to protect BCMEC against apoptosis induced by hydrogen peroxide. CONCLUSION: Hydrogen peroxide induces BCMEC injury either by apoptosis or through necrosis. Hydroxyethylpuerarin protects BCMEC against hydrogen peroxide-induced injury in a concentration-dependent manner. Its antioxidant effects might be involved as the mechanism protection.

Publication Types: PMID: 15952592 [PubMed - indexed for MEDLINE]

152. Life-threatening interaction between the root extract of Pueraria lobata and methotrexate in rats.

Chiang HM, Fang SH, Wen KC, Hsiu SL, Tsai SY, Hou YC, Chi YC, Chao PD.

Institute of Pharmaceutical Chemistry, China Medical University, Taichung, Taiwan, ROC.

Isoflavone supplements are nowadays widely used as alternative for hormone replacement therapy. However, the safety remains unanswered. This study attempted to investigate the effect of Pueraria lobata root decoction (PLRD), an isoflavone-rich herb, on the pharmacokinetics of methotrexate (MTX), a bicarboxylate antimetabolite with narrow therapeutic window. Rats were orally and intravenously given methotrexate alone and coadministered with PLRD. Blood samples were withdrawn via cardiopuncture at specific time points after drug administration. Serum methotrexate concentrations were assayed by specific monoclonal fluorescence polarization immunoassay method. Pharmacokinetic parameters were calculated using noncompartment model of WINNONLIN for both oral and intravenous data of MTX. Our results showed that coadministration of 4.0 g/kg and 2.0 g/kg of PLRD significantly increased the AUC(0-t) by 207.8% and 127.9%, prolonged the mean residence time (MRT) by 237.8 and 155.2%, respectively, finally resulted in surprisingly high mortalities of 57.1% and 14.3% in rats. When MTX was given intravenously, the coadministration of PLRD at 4.0 g/kg significantly increased the half-life by 53.9% and decreased the clearance by 47.9%. In conclusion, the coadministration of PLRD significantly decreased the elimination and resulted in markedly increased exposure of MTX in rats.

Publication Types: PMID: 15936791 [PubMed - indexed for MEDLINE]

153. A glycoside flavonoid in Kudzu (Pueraria lobata): identification, quantification, and determination of antioxidant activity.

Lau CS, Carrier DJ, Beitle RR, Howard LR, Lay JO, Liyanage R, Clausen EC.

Department of Chemical Engineering, 3202 Bell Engineering Center, University of Arkansas, 115 Chemistry Building, Fayetteville, AR 72701, USA.

Kudzu (Pueraria lobata) foliage has been touted as a possible energy crop. High-performance liquid chromatography and mass spectrometry analysis of the methanolic kudzu foliage extracts confirmed the presence of robinin (kaempferol-3-O-robinoside-7-O-rhamnoside). Robinin accounted for 0.65 +/- 0.16% (dry basis) of kudzu biomass. Fast performance liquid chromatography (FPLC) was employed to fractionate robinin from the crude extract. The antioxidant capacity of robinin was evaluated by an oxygen radical absorbance capacity (ORAC) assay. The ORAC values of pure standard were compared with those of the extract fractions. One milligram of the FPLC-fractionated robinin generated an ORAC value of 5.15 +/- 2.00 micromol/mg of Trolox, whereas 1 mg of pure robinin generated an ORAC value of 12.34 +/- 0.45 micromol/mg of Trolox. Because of its antioxidant properties, robinin may be a flavonoid worth extracting prior to energy production.

Publication Types: PMID: 15930558 [PubMed - indexed for MEDLINE]

154. An extract of the Chinese herbal root kudzu reduces alcohol drinking by heavy drinkers in a naturalistic setting.

Lukas SE, Penetar D, Berko J, Vicens L, Palmer C, Mallya G, Macklin EA, Lee DY.

Behavioral Psychopharmacology Research Laboratory, McLean Hospital, Belmont, Massachusetts 02478, USA. lukas@mclean.harvard.edu

BACKGROUND: Of the available medications for treating alcohol-related problems, none are universally effective, and all have side effects that may limit their use. Extracts of kudzu containing a variety of isoflavones have been shown to reduce alcohol drinking in rats and hamsters. METHODS: The present study was designed to test the efficacy of a kudzu extract in a clinical population. Male and female "heavy" alcohol drinkers were treated with either placebo or a kudzu extract for 7 days and then given an opportunity to drink their preferred brand of beer while in a naturalistic laboratory setting. Participants served as their own controls, and order of treatment exposure was counterbalanced. Drinking behavior was monitored by a digital scale that was located in the top of an end table. RESULTS: Kudzu treatment resulted in significant reduction in the number of beers consumed that was paralleled by an increase in the number of sips and the time to consume each beer and a decrease in the volume of each sip. These changes occurred in the absence of a significant effect on the urge to drink alcohol. There were no reported side effects of kudzu treatment. CONCLUSION: These data suggest that an extract of this leguminous plant may be a useful adjunct in reducing alcohol intake in a naturalistic setting.

Publication Types: PMID: 15897719 [PubMed - indexed for MEDLINE]

155. Antitumor activity of spinasterol isolated from Pueraria roots.

Jeon GC, Park MS, Yoon DY, Shin CH, Sin HS, Um SJ.

Department of Bioscience and Biotechnology/Institute of Bioscience, Sejong University, Seoul 143-747, Korea.

We purified phytoestrogens from Pueraria root (Pueraria mirifica from Thailand and Pueraria lobata from Korea), which is used as a rejuvenating folk medicine in Thailand and China. Dried, powdered plant material was extracted with 100% ethanol and further separated by concentration, filtration, and thin layer silica gel chromatography. Using the fractions obtained during separation, we first investigated their cytotoxicity in several cancer cell lines from various tissues. The ethanol-extracted components (PE1, PE4) had significant antiproliferative effects on breast cancer cell lines, including MCF-7, ZR-75-1, MDA-MB-231, SK-BR-3, and Hs578T. Second, we compared these results with the cytotoxic effects of known flavonoids, sterols, and coumarins from Pueraria root. The known compounds were not as effective, and occurred in a different polarity region on HPLC. Third, further separation resulted in the isolation of eight different components (Sub PE-A to -H). One of these, PE-D, affected the growth of some breast cancer cell lines (MCF-7, MDA- MB-231) in a dose- and time-dependent manner, as well as the growth of ovarian (2774) and cervical cancer cells (HeLa). Finally, a transfection assay showed that this component had an estrogenic effect similar to 17beta - estradiol, which activates both estrogen receptor alpha (ERalpha) and ERbeta. The NMR analysis determined that spinasterol (stigmasta-7, 22-dien-3beta-ol) is an active cytotoxic component of Pueraria root.

Publication Types: PMID: 15886524 [PubMed - indexed for MEDLINE]

156. Analysis of the estrogenic components in kudzu root by bioassay and high performance liquid chromatography.

Zhang Y, Chen J, Zhang C, Wu W, Liang X.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 161 Zhongshan Road, Dalian 116011, PR China.

The estrogenic activity of the Chinese herb kudzu root was investigated by a recombinant yeast screening assay (YES). Isoflavones are the main components in the plant, of which puerarin is the most abundant one. The kudzu root extract was separated into four fractions according to the polarity. The crude extract and its sub-fractions, except the water fraction, showed clear estrogenic activity and the potencies were in the range of 10(-3) to 10(-1)g/l. The ligand potency was used to compare the estrogenic activity of these fractions. The crude extract and its sub-fractions were further analyzed by high performance liquid chromatography (HPLC) to correlate the activity and the active components. Bioassay and chemical analysis showed that theoretical estrogenic activity expressed as equivalent 17beta-estradiol concentration or the cumulative effects are comparable to that experimentally determined by YES. The results showed that the high content of isoflavones as well as the high estrogenic activity could make kudzu root extract an interesting candidate for hormone replacement therapy.

Publication Types: PMID: 15857757 [PubMed - indexed for MEDLINE]

157. Kinetic modeling of the adsorption of basic dyes by kudzu.

Allen SJ, Gan Q, Matthews R, Johnson PA.

School of Chemical Engineering, Queen's University of Belfast, Stranmillis Road, Belfast BT9 5AG, UK. s.allen@qub.ac.uk

The use of kudzu, a rapidly growing, high-climbing perennial leguminous vine, for the adsorption of basic dyes from aqueous solution has been investigated at various initial dye concentrations, masses of kudzu, and agitation rates. The extent and rate of adsorption of the three basic dyes (Basic Red 22, Basic Yellow 21, and Basic Blue 3) were analyzed using a pseudo-first-order and a pseudo-second-order kinetic model. While both rate mechanisms provided an acceptable degree of correlation with the experimental sorption rate data, the pseudo-second-order model gave a much higher degree of correlation, suggesting that this model could be used in design and simulation applications.

PMID: 15848407 [PubMed - indexed for MEDLINE]

158. In vitro estrogenic activities of Chinese medicinal plants traditionally used for the management of menopausal symptoms.

Zhang CZ, Wang SX, Zhang Y, Chen JP, Liang XM.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Zhongshan Road No. 161, Dalian 116011, PR China.

The estrogenic activity of 70% EtOH extracts of 32 traditional Chinese medicinal plants, selected according to their reported efficacy for the treatment of menopausal symptoms, was assessed using a recombinant yeast system with both a human estrogen receptor expression plasmid and a reporter plasmid. Among them, 11 (34%) species proved to be active. Polygonum cuspidatum had the highest estrogenic relative potency (RP) (3.28 x 10(-3)), followed by Rheumpalmatum (3.85 x 10(-4)), Cassia obtusifolia (3.49 x 10(-4)), Polygonum multiflorum (2.87 x 10(-4)), Epimedium brevicornum (2.30 x 10(-4)), Psoralea corylifolia (1.90 x 10(-4)), Cynomorium songaricum (1.78 x 10(-4)), Belamcanda chinensis (1.26 x 10(-4)), Scutellaria baicalensis (8.77 x 10(-5)), Astragalus membranaceus (8.47 x 10(-5)) and Pueraria lobata (6.17 x 10(-5)). The EC(50) value of 17beta-estradiol used as the positive control was 0.205+/-0.025 ng/ml (RP=100). This study gave support to the reported efficacy of Chinese medicines used for hormone replacement therapy.

Publication Types: PMID: 15814262 [PubMed - indexed for MEDLINE]

159. Ovulation block by Pueraria mirifica: a study of its endocrinological effect in female monkeys.

Trisomboon H, Malaivijitnond S, Watanabe G, Taya K.

Department of Physiology, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand.

Pueraria mirifica (PM), a Thai herb containing phytoestrogens, may act as estrogen and disturb reproduction. To investigate the effect of PM on the menstrual cycle length and related hormones, nine adult female monkeys (Macaca fascicularis) were separated into three groups. Each group (n = 3) was fed with 10, 100, and 1000 mg/d of PM for three menstrual cycles. The menstrual cycle length increased significantly in monkeys treated with PM-10 and PM-100 and disappeared completely in monkeys treated with PM-1000. Serum follicle stimulating hormone, luteinizing hormone, estradiol, progesterone, and irinhibin were lower during the treatment period in a dose-dependent manner. Changes in menstrual cycle length and the hormonal levels recovered during the post-treatment period only in monkeys treated with PM-10 and PM-100. PM greatly influences menstrual cycles and may suppress ovulation by lowering serum levels of gonadotropins.

PMID: 15805583 [PubMed - indexed for MEDLINE]

160. Efficient production of (2S)-flavanones by Escherichia coli containing an artificial biosynthetic gene cluster.

Miyahisa I, Kaneko M, Funa N, Kawasaki H, Kojima H, Ohnishi Y, Horinouchi S.

Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan. aa36248@mail.ecc.u-tokyo.ac.jp

For the fermentative production of plant-specific flavanones (naringenin, pinocembrin) by Escherichia coli, a plasmid was constructed which carried an artificial biosynthetic gene cluster, including PAL encoding a phenylalanine ammonia-lyase from a yeast, ScCCL encoding a cinnamate/coumarate:CoA ligase from the actinomycete Streptomyces coelicolor A3(2), CHS encoding a chalcone synthase from a licorice plant and CHI encoding a chalcone isomerase from the Pueraria plant. The recombinant E. coli cells produced (2S)-naringenin from tyrosine and (2S)-pinocembrin from phenylalanine. When the two subunit genes of acetyl-CoA carboxylase from Corynebacterium glutamicum were expressed under the control of the T7 promoter and the ribosome-binding sequence in the recombinant E. coli cells, the flavanone yields were greatly increased, probably because enhanced expression of acetyl-CoA carboxylase increased a pool of malonyl-CoA that was available for flavanone synthesis. Under cultural conditions where E. coli at a cell density of 50 g/l was incubated in the presence of 3 mM tyrosine or phenylalanine, the yields of naringenin and pinocembrin reached about 60 mg/l. The fermentative production of flavanones in E. coli is the first step in the construction of a library of flavonoid compounds and un-natural flavonoids in bacteria.

Publication Types: PMID: 15770480 [PubMed - indexed for MEDLINE]

161. Evaluation of the estrogenic activity of Leguminosae plants.

Yoo HH, Kim T, Ahn S, Kim YJ, Kim HY, Piao XL, Park JH.

College of Pharmacy, Seoul National University, Seoul, Korea.

The plant extracts of the Leguminosae family were screened for their estrogenic activity with the Ishikawa cell system. Of the tested plants, Desmodium oxyphyllum, Dunbaria villosa, Kummerowia striata, Lespedeza bicolor, Maackia amurensis, Maackia fauriei, Pueraria thunbergiana, and Sophora flavescens were highly estrogenic with EC50 values of less than 10 microg/ml.

Publication Types: PMID: 15744086 [PubMed - indexed for MEDLINE]

162. Hepatoprotective effects of irisolidone on tert-butyl hyperoxide-induced liver injury.

Lee HU, Bae EA, Kim DH.

Kyung Hee University, College of Pharmacy, Hoegi, Dongdaemun-ku, Seoul, Korea.

To clarify the hepatoprotective effects of kakkalide and its metabolite irisolidone by human fecal microflora, their effects on tert-butyl hydroperoxide (t-BHP)-injured HepG2 cells and mice were investigated. Irisolidone protected HepG2 cells against cytotoxicity induced by t-BHP. However, kakkalide did not protect cytotoxicity. When kakkalide 100 mg/kg was orally administered to mice injured by t-BHP, it significantly inhibited the increase in plasma alanine aminotransferase and aspartate aminotransferase activities by 84% and 85% of t-BHP-treated control group, respectively. The inhibitory effect of kakkalide is much more potent than that of silybin, a hepatoprotective agent. However, intraperitoneally administered kakkalide did not exhibit hepatoprotective activity. When irisolidone was intraperitoneally administered to mice, it exhibited potent hepatoprotective activity. Based on these findings, irisolidone can be hepatoprotective and kakkalide may be a prodrug transformed to irisolidone.

Publication Types: PMID: 15744084 [PubMed - indexed for MEDLINE]

163. [Effect of shenwu capsule on the peripheral blood lymphocyte DNA of rat model induced by beta-amyloid injection]

[Article in Chinese]

Wang H, Li L, Ye CF, Zhang L.

Medical Sciences, Beijing Key Laboratory for Brain Aging, Beijing, 100053, China.

OBJECTIVE: To investigate the effect of the new Traditional Chinese Compound Shenwu Capsule on the damage of lymphocyte DNA and lipid peroxidation in peripheral blood of rats induced by beta-amyloid injection. METHOD: The animal model was made by injection of beta-amyloid25-35 into hippocampus of rats. DNA damage of lymphocytes was measured by the single cell gel electrophoresis (SCGE) combined with the laser scanning confocal microscopy (LSCM). The content of malondialdehyde (MDA) was determined by TBA assay. RESULT: Shenwu Capsule decreased the rate of the comet-like cell, comet-like cell lengh (TCL), and tail moment (TM) of lymphocytes in peripheral blood of model rats. Shenwu Capsule also declined the MDA content in serum of Abeta model rats. CONCLUSION: Shenwu Capsule has protective effect on peripheral blood lymphocyte DNA.

Publication Types: PMID: 15719690 [PubMed - indexed for MEDLINE]

164. [Effects of puerarin on plasma membrane GLUT4 content in skeletal muscle from insulin-resistant Sprague-Dawley rats under insulin stimulation]

[Article in Chinese]

Song CY, Bi HM.

Department of Endocrinology, Renmin Hospital of Wuhan University, Wuhan 430060, China. lynn-song@163.com

OBJECTIVE: To explore the effect of puerarin injection on the amount of GLUT4 protein at the plasma membrane in insulin-resistant rat skeletal muscle. METHOD: The rat model of insulin resistance (IR) was made by being fed with high-fat diet. The animals were divided into three groups (ten in each group): group I: controls; group II: Insulin-resistant rats; group III: Insulin-resistant rats + Puerarin treatment. Insulin-resistant rats were injected with 100 mg puerarin injection per kg body weight through abdominal cavity once a day for 4 weeks. Fasting blood glucose and fasting serum insulin levels were measured before and after Puerarin treatment, respectively. Insulin treatment was achieved by intraperitoneal injection of insulin (1 unit insulin per kg body weight.) 15 minute before killing the animals. The right hindlimb skeletal muscle was rapidly dissected. Then the expression of GLUT4 protein at the plasma membrane in all the animals was assessed with Western blot method. RESULT: The GLUT4 content at the plasma membrane in insulin-resistant rats skeletal muscle was significantly lower (about 31%) than that of controls (P < 0.01). Puerarin Injection partly corrected fasting blood glucose (from 6.17 +/- 0.67 mmol x L(-1) to 5.54 +/- 0.35 mmol x L(-1)) and fasting serum insulin levels (from 17.09 +/- 2.02 mU x L(-1) to 11.86 +/- 1.35 mU x L(-1)) and increased the GLUT4 content at the plasma membrane by 1.18-fold in insulin-resistant rats skeletal muscle. CONCLUSION: Puerarin Injection can ameliorate IR, and the mechanism may be involved in increasing cell-surface level of GLUT4 through decreasing fasting blood glucose and fasting serum insulin levels, improving GLUT4 trafficking and intracellular insulin signaling.

Publication Types: PMID: 15719689 [PubMed - indexed for MEDLINE]

165. [Neuro-protective effect of naomaitong to brain damage after focal cerebral ischemia reperfusion (I/R) in the aged rats]

[Article in Chinese]

Ren XQ, Li JS, Feng YM, Lu YQ.

Henan College of Traditional Chinese Medicine, Zhengzhou 450008, China. Rxq23@sohu.com

OBJECTIVE: To study neuro-protective effect of naomaitong on brain damage after focal cerebral ischemia reperfusion (I/R) in the aged rats. METHOD: 11 groups of 20-22-month old SD rats were subjected to 3 hours of middle cerebral artery occlusion with the intraluminal filament technique, followed by 12 hours of reperfusion. Nervous symptom, oedema of brain, infarct size, morphology and superfine structure of brain, etc were monitored thoughout cerebral ischemia and reperfusion. RESULT: Infarct size of brain in ischemia and reperfusion groups were significantly greater. Oedema of it was higher, nervous symptom of it was more serious, and morphology and superfine structure brain were more obvious than those of sham-operated group. Nervous symptom, oedema of brain, infarct size, morphology and superfine structure of brain in treated groups were ligher than those of untreated groups (P < 0.001). CONCLUSION: Naomaitong can protect brain damage after focal cerebral I/R in the aged rats.

Publication Types: PMID: 15709387 [PubMed - indexed for MEDLINE]
166. HPLC method for the determination and pharmacokinetic studies on puerarin in cerebral ischemia reperfusion rat plasma after intravenous administration of puerariae radix isoflavone.

Yan B, Xing D, Ding Y, Tao J, Du LJ.

Institute of Medicinal Plant, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100094, China.

A new HPLC method for the determination of puerarin in cerebral ischemia reperfusion rat plasma is introduced. Puerarin, the principal bioactive component of puerariae radix isoflavone, was extracted from plasma by methanol. The HPLC separation was then performed on a reversed-phase C18 column using water-acetonitrile (89:11, v/v) as eluting solvent system, and UV detection at 252 nm to measure the analyte with a limit of quantitation about 9.44 ng ml(-1). The calibration curve for puerarin was linear (r=0.9998) in the concentration range of 9.44-1208.00 ng ml(-1), both intra- and inter-day precision of the puerarin were determined and their coefficient of variation did not exceed 10%. The validated method has been successfully applied for pharmacokinetic studies of puerarin from rat plasma after intravenous administration of puerariae radix isoflavone. Another novel finding of this study was that the elimination rate of puerarin was significantly slower in the cerebral ischemia reperfusion rat than in the normal rat, judging by the pharmacokinetic parameters obtained. Since puerariae radix isoflavone was mainly administrated to the patients suffering from cerebralvascular diseases, the pharmacokinetic studies performed on the pathological animal models were suitable references for clinical application.

PMID: 15708670 [PubMed - indexed for MEDLINE]

167. [Effect of puerarin and daidzein on proliferating vascular smooth muscle cells]

[Article in Chinese]

Han J, Wang W, Wang LY, Liu S, Kang TD.

Dongzhimen Hospital, University of Traditional Chinese Medicine, Beijing 100029, China.

OBJECTIVE: To observe the different effects of Puerarin and Daidzein on the expression of proliferating vascular smooth muscle cells, and to discuss the mechanism. METHOD: MT was used to detect the state of VSMC (vascular smooth muscle cell) activity. The expression levels of Survivin, Bcl-xl, Bax and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) messenger RNA (mRNA) were analyzed quantitatively by reverse transcriptase polymerase chain reaction (Rt-PCR). RESULT: Compared with Puerarin groups, VSMC activity in daidzein groups was lower, and the ratio of Bax/Gapdh/Bcl-xl/Gapdh was higher. CONCLUSION: The inhibition effect of daidzein on VSMC proliferation is stronger than that of puerarin.

Publication Types: PMID: 15706898 [PubMed - indexed for MEDLINE]

168. [Influence of Pueraria thomsonii on insulin resistance induced by dexamethasone]

[Article in Chinese]

Bai HY, Zou WJ, Gao XP.

Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.

OBJECTIVE: To investigate the influence of Pueraria thomsonii on insulin resistance induced by dexamethasone. METHOD: 3T3-L1 adipocytes model of insulin resistance was made by dexamethasone and the change of glucose concentration in cell culture was determined after action of drugs. Rat animal model of insulin resistance was made by intramuscular dexamethasone (1 mg x kg(-1), every other day), and fasting blood glucose (FBG) and fasting serum insulin (FINS) were enamined, and at the end of the experiment, insulin sensitive index (ISI) and insulin resistance index (IRI) were calculated. RESULT: P. thomsonii decreased the concentration of glucose in 3T3-L1 adipocytes culture significantly and improved the sensitivity of 3T3-L1 adipocytes to insulin. P. thomsonii improved the sensitivity of rats to insulin and diminished the fasting serum insulin and insulin resistance index. CONCLUSION: P. thomsonii can significantly improve insulin resistance induced by dexamethasone.

Publication Types: PMID: 15706877 [PubMed - indexed for MEDLINE]

169. [Effect of puerarin on L-type calcium channel in isolated rat ventricular myocytes]

[Article in Chinese]

Guo XG, Chen JZ, Zhang X, Xia Q.

Department of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China. gxg@mail.hz.zj.cn

OBJECTIVE: To observe the effect of Puerarin on L-type calcium channel in isolated rat ventricular myocytes. METHOD: The cardiac ventricular myocytes were isolated enzymatically by Langendorff perfusion techniques at constant flow rate. Whole-cell recording of patch-clamp techniques was used to observe the current of L-type calcium channel. RESULT: Puerarin 2.4 mmol x L(-1) could inhibit the current of L-type calcium channel of rat ventricular myocytes and this inhibition was time-dependent. Purerarin elevated the current-voltage (I-V) curve of calcium current. CONCLUSION: Puerarin can inhibit L-type calcium current of rat ventricular myocytes. Which implies that puerarin takes part in anti-myocardial ischemia and anti-arrhythmics partly due to the inhibition of L-type calcium channel.

Publication Types: PMID: 15706853 [PubMed - indexed for MEDLINE]

170.The first total synthesis of kwakhurin, a characteristic component of a rejuvenating plant, "kwao keur": toward an efficient synthetic route to phytoestrogenic isoflavones.

Ito F, Iwasaki M, Watanabe T, Ishikawa T, Higuchi Y.

Graduate School of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi, Inage, Chiba 263-8522, Japan.

A convergent synthesis of kwakhurin (5), a characteristic estrogen-like isoflavone of Pueraria mirifica(Leguminosae), is described. Isoflavone skeleton 31 was constructed by Suzuki-Miyaura coupling of 3-bromochromone 26 (AC-ring) and arylboronic acid 30 (B-ring) in the presence of TBAB as an additive. Microwave-assisted coupling was also examined, but did not improve the yield. Baeyer-Villiger oxidation, followed by propargylation and reduction afforded 1,1-dimethylallyl ether 37. 6'-Prenylisoflavone 34 was obtained in high yield by Claisen rearrangement of 37 in N,N-diethylaniline. On the other hand, 1,3-rearrangement of prenyl ether 33 with clay gave 34 in poor yield. Successive methylation of 34 and deprotection yielded the target kwakhurin (5) in 12% overall yield from 2,4-dihydroxybenzaldehyde (23).

PMID: 15703807 [PubMed - indexed for MEDLINE]

171. Analysis of isoflavone daidzein in Puerariae radix with micelle-mediated extraction and preconcentration.

He J, Zhao Z, Shi Z, Zhao M, Li Y, Chang W.

Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, People's Republic of China.

Nonionic surfactant oligo(ethylene glycol) monoalkyl ether (Genapol X-080) was employed as an alternative and effective solvent for the extraction of daidzein from Puerariae radix for the first time. Optimum experimental conditions were established. With 5% Genapol X-080 (w/v), liquid/solid ratio of 25:1 (mL/g), and ultrasonic-assisted extraction for 45 min, the extraction percentage of daidzein reached the highest value. For the preconcentration of daidzein by cloud-point extraction (CPE), sodium chloride was added to the solution to facilitate the phase separation and increase the preconcentration factor by reducing the volume of the surfactant-rich phase. The preconcentration factor for daidzein was about 13. Satisfactory results were obtained for the analysis of daidzein from P. radix with this established method.

Publication Types: PMID: 15686396 [PubMed - indexed for MEDLINE]

172. Evolution of the isoprene biosynthetic pathway in kudzu.

Sharkey TD, Yeh S, Wiberley AE, Falbel TG, Gong D, Fernandez DE.


Department of Botany, University of Wisconsin, Madison, Wisconsin 53706, USA. tsharkey@wisc.edu

Isoprene synthase converts dimethylallyl diphosphate, derived from the methylerythritol 4-phosphate (MEP) pathway, to isoprene. Isoprene is made by some plants in substantial amounts, which affects atmospheric chemistry, while other plants make no isoprene. As part of our long-term study of isoprene synthesis, the genetics of the isoprene biosynthetic pathway of the isoprene emitter, kudzu (Pueraria montana), was compared with similar genes in Arabidopsis (Arabidopsis thaliana), which does not make isoprene. The MEP pathway genes in kudzu were similar to the corresponding Arabidopsis genes. Isoprene synthase genes of kudzu and aspen (Populus tremuloides) were cloned to compare their divergence with the divergence seen in MEP pathway genes. Phylogenetic analysis of the terpene synthase gene family indicated that isoprene synthases are either within the monoterpene synthase clade or sister to it. In Arabidopsis, the gene most similar to isoprene synthase is a myrcene/ocimene (acyclic monoterpenes) synthase. Two phenylalanine residues found exclusively in isoprene synthases make the active site smaller than other terpene synthase enzymes, possibly conferring specificity for the five-carbon substrate rather than precursors of the larger isoprenoids. Expression of the kudzu isoprene synthase gene in Arabidopsis caused Arabidopsis to emit isoprene, indicating that whether or not a plant emits isoprene depends on whether or not it has a terpene synthase capable of using dimethylallyl diphosphate.

Publication Types: PMID: 15653811 [PubMed - indexed for MEDLINE] PMCID: PMC1065370

173. Long-term treatment effects of Pueraria mirifica phytoestrogens on parathyroid hormone and calcium levels in aged menopausal cynomolgus monkeys.

Trisomboon H, Malaivijitnond S, Suzuki J, Hamada Y, Watanabe G, Taya K.

Biological Science Ph.D. Program, Faculty of Science, Chulalongkorn University, Bangkok, Tokyo, Japan.

To determine the effect of Pueraria mirifica (PM) on serum parathyroid hormone (PTH) and calcium levels on aged menopausal monkeys (Macaca fascicularis), subjects were treated with 10, 100, or 1,000 mg/day of PM. Blood samples were collected every 5 days for 30, 90, and 60 days during pre-treatment, treatment, and post-treatment periods, respectively. Sera were assayed for PTH, estradiol, and calcium levels. PM-1,000 had the strongest effect on the decrease in PTH (0.001<P< or =0.05) and calcium levels (0.001<P< or =0.03) during the treatment period. PTH levels remained low for the first 15 days of the post-treatment period (0.01< or =P < or =0.05). PM-10 induced a significant decrease in PTH level on day 80 (P=0.02) during the treatment period and a significant decrease in calcium level on day 75 (P<0.01). There were no changes in serum PTH and calcium levels throughout the study period in the PM-100 group. Estradiol levels decreased significantly during the treatment period in all treatment groups. The results suggest that long-term treatment with 1,000 mg/day of PM decreases serum PTH and calcium levels in aged menopausal monkeys, indicating that PM ameliorates bone loss caused by estrogen deficiency.

Publication Types: PMID: 15647615 [PubMed - indexed for MEDLINE]

174. [Effect of sucrose concentration on the growth and production of secondary metabolites in Pueraria phaseoloides hairy roots]

[Article in Chinese]

Liang P, Shi HP, Qi Y.

Guangdong Key Lab of Biotechnology for Plant Development, College of Life Science, South China Normal University, Guangzhou 510631, China.

Effects of sucrose concentrations on the growth and production of puerarin and isoflavones compounds in Pueraria phaseoloides hairy roots induced by Agrobacterium rhizogenes ATCC15834 were investigated. Changes of sucrose consumption in the medium during liquid culture were also determined. The results showed cultured for 16 days in MS medium with 5%, 4%, 3% and 2% sucrose, the proliferation times of dry weight of hairy roots were 11.7, 11.9, 10.1 and 5.9, respectively. 3% sucrose concentration in liquid medium was the best for accumulation of puerarin and isoflavones in the hairy roots. The highest content of puerarin, 5.147 mg/g DW, was obtained after 12 days of liquid culturing while the highest content of isoflavones, about 27.76 mg.g.DW, was gained after 16 days in culturing. Sucrose concentration decreased as hairy root growth proceeded. The growth rate and the content of soluble sugar in hairy roots of P. phaseoloides was directly proportional to the rate of sucrose utilization in the liquid medium during the whole culture. It was observed that the highest content of soluble sugar in hairy roots was at day 12 of liquid culture and sucrose in the liquid medium was used up at the end of 16 days of culture.

Publication Types: PMID: 15636366 [PubMed - in process]

175. [Endothelium-independent vasorelaxant effect of puerarin on rat thoracic aorta]

[Article in Chinese]

Dong K, Tao QM, Xia Q, Shan QX, Pan GB.

Department of Cardiovascular, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China. dongkan76@sohu.com

OBJECTIVE: To investigate the vasorelaxant effect of puerarin in rat aortic rings and the mechanism. METHOD: The isolated thoracic aortic rings of male Sprague-Dawley rats were mounted on the organ bath and the contractile responses of the vessel were recorded. RESULT: Puerarin completely relaxed the contractions induced by phenylephrine in a concentration-dependent manner in endothelium-intact and endothelium-denuded rat aorta, but it had no effect on those preconstricted by a high concentration of potassium chloride (KCl, 60 mmol x L(-1)). The relaxant effect of puerarin was significantly inhibited by pretreatment of endothelium-denuded aorta with potassium channel antagonists tetraethylammonium, 4-aminopyridine but not glibenclamide. CONCLUSION: Puerarin induces an endothelium-independent relaxation in rat aortic rings. The mechanisms may involve the reduction in Ca2+ influx through the calcium channels operated by alpha-adrenergic receptor and the activation of the potassium channels (Kv and BKca, but not KATP).

Publication Types: PMID: 15631089 [PubMed - indexed for MEDLINE]

176. Study on the bioavailability of puerarin from Pueraria lobata isoflavone self-microemulsifying drug-delivery systems and tablets in rabbits by liquid chromatography-mass spectrometry.

Cui S, Zhao C, Tang X, Chen D, He Z.

School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, People's Republic of China.

To evaluate the bioavailability of puerarin from Pueraria lobata isoflavone self-microemulsifying drug delivery systems (SMEDDS) and Yufengningxin tablets, a rapid and specific liquid chromatography--mass spectrometric method was developed and validated to determine puerarin in rabbit serum. The analyte was extracted from serum samples by precipitating the serum proteins, separated on a Diamonsil C(18) column and detected by mass spectrometry with an electrospray ionization interface. 4-Hydroxybenzaldehyde was used as the internal standard. The method has a limit of quantitation of 10 ng/mL using 200 microL serum. The intra-day relative standard deviations (RSDs) ranged from 3.7 to 6.9% and inter-day RSDs were within 6.5%. After administration of SMEDDS and tablets to rabbits, a significant difference was observed in main pharmacokinetic parameters of t(max), C(max) and AUC(0--infinity) between SMEDDS and tablets, and a 2.2-fold increase in the relative bioavailability of puerarin was observed with the SMEDDS compared with Yufengningxin tablets. It was concluded that the absorption of puerarin from Pueraria lobata isoflavone SMEDDS was enhanced. (c) 2004 John Wiley & Sons, Ltd.

Publication Types: PMID: 15627278 [PubMed - indexed for MEDLINE]

177. [Experimental study of puerarin injection on the hemorheology in acute blood-stasis model rats]

[Article in Chinese]

Pan HP, Yang JZ, Li LL, Yi F, Huang ZQ, Huang KW.

People's Hospital of Guangxi Autonomous Region, Nanning 530021, Guangxi, China.

OBJECTIVE: To observe the effect of Puerarin Injection on the hemorheology in acute blood-stasis model rats. METHOD: The acute blood-stasis model rats were made by being soaked in ice water afer being injected adrenaline hydrochloride injection in a major dose. The changes of viscosity of whole blood and plasma, blood yield stress, erythrocyte aggregation and the maximum rate of platelet aggregation in the acute blood-stasis model rats were measured with Auto-Viscometer, and then the influence of Puerarin Injection on the hemorheology in the model rats was investigated. RESULT: The viscosity of whole blood and plasma, and blood yield stress in the acute blood-stasis model rats were significantly higher than those in the normal control group (P < 0.01, P < 0.05). Both the high dose and the low dose of Puerarin Injection could reduce the viscosity of whole blood and plasma, blood yield stress and the maximum rate of platelet aggregation in the acute blood-stasis model rats (P < 0.01, P < 0.05). The high dose could also reduce the erythrocyte aggregation and the deformed Index of red blood cell (P < 0.05). CONCLUSION: Puerarin Injection can ameliorate the hemorheology in acute blood-stasis model rats, and it has a dose-response relationship.

Publication Types: PMID: 15617506 [PubMed - indexed for MEDLINE]

178. [Study on HPLC-FPS of Pueraria of different sources]

[Article in Chinese]

He CN, Li M, Cao ZG, Guo HY, Wang CL, Yu SC.

Institute of Medicinal Plant, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100094, China. chunnianhe@163.com

OBJECTIVE: To make comparative study on HPLC-FPS of several kinds of Pueraria lobata and P. thomsonii from different sources. METHOD: Kromasil C18 column was used, with mixture of acetonitrile and water as mobile phase in a gradient mode. The wavelength of measurement was 250 nm. RESULT AND CONCLUSION: The fingerprints of P. lobata and P. thomsonii were obtained. This method can be used to identify P. lobata and P. thomsonii from different sources conveniently, and it may be practically valuable for the quality control of sample for P. lobata or P. thomsonii and its preparation.

Publication Types: PMID: 15617494 [PubMed - indexed for MEDLINE]

179. Puerarin acts through brain serotonergic mechanisms to induce thermal effects.

Chueh FS, Chang CP, Chio CC, Lin MT.

Department of Applied Life Science, Taichung Healthcare and Management University, Taichung, Taiwan 413.

The present study was attempted to investigate the effect of puerarin, an isoflavone compound isolated from Pueraria lobata, on both the basal body temperature and pyrogenic fever in unanesthetized, restrained rats. Intraperitoneal administration of puerarin or crude extracts of Pueraria lobata elicited hypothermia. Direct administration of a small amount of puerarin into the lateral cerebral ventricle produced the same extent of hypothermia. Systemic or central administration of puerarin causes a decrease in both colonic temperature and hypothalamic 5-HT efflux in rats. The puerarin-induced hypothermia and decreased 5-HT efflux in the hypothalamus were attenuated by selective depletion of hypothalamic 5-HT produced by intracerebroventricular injection of 5,7-dihydroxytryptamine. Furthermore, the puerarin-induced hypothermia was almost completely abolished by treatment with a 5-HT2A-receptor agonist (DOI or quipazine) or a 5-HT1A-receptor antagonist [(-)-pindolol]. A 5-HT2A-receptor antagonist (ketanserin) or a 5-HT1A-receptor agonist (8-OH-DPAT) had additive effects with puerarin. Intracerebroventricular administration of interleukin-1 caused an increase in both colonic temperature and hypothalamic 5-HT efflux. The interleukin-1-induced hyperthermia and increased 5-HT efflux in the hypothalamus were attenuated by treatment with systemic administration of puerarin. The data indicate that puerarin exerts its hypothermic and antipyretic effects by activating 5-HT1 receptor and/or antagonizing 5-HT2A receptors in the hypothalamus.

Publication Types: PMID: 15599109 [PubMed - indexed for MEDLINE]

180. Different effects of Pueraria mirifica, a herb containing phytoestrogens, on LH and FSH secretion in gonadectomized female and male rats.

Malaivijitnond S, Kiatthaipipat P, Cherdshewasart W, Watanabe G, Taya K.

Primate Research Unit, Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand. Suchinda.m@chula.ac.th

To investigate the effect of Pueraria mirifica (P. mirifica) containing phytoestrogens on reproductive systems, both sexes of rats were gonadectomized and treated orally with 0, 10, 100, and 1,000 mg/kg BW per day of P. mirifica suspended in water (abbreviated as P-0, P-10, P-100, and P-1000), respectively. The treatment schedule was separated into 3 periods: pre-treatment, treatment, and post-treatment. The duration for each period was 14 days. Blood samples were taken once a week. Serum LH and FSH levels were significantly increased within 1 week after gonadectomy; and there were no changes after administration of P-0, P-10, and P-100. However, the increase of LH levels in both sexes and FSH levels in females were attenuated within 1 week after P-1,000 treatment. The attenuation of LH levels in males was smaller than that of females. The decrease of gonadotropin levels was recovered within 1 week in males and 2 weeks in females, respectively, during the post-treatment period. The increase of uterine weight and vaginal cornification were observed in female rats treated with P-100 and P-1,000, whereas only the increase of epididymis weight was found in male rats treated with P-1,000. From this study, it can be concluded that P. mirifica can influence the reproductive functions in both sexes of rats, but the response in females is greater than in males.

Publication Types: PMID: 15599108 [PubMed - indexed for MEDLINE]

181. A comparative study on aqueous root extracts of Pueraria thomsonii and Pueraria lobata by antioxidant assay and HPLC fingerprint analysis.

Jiang RW, Lau KM, Lam HM, Yam WS, Leung LK, Choi KL, Waye MM, Mak TC, Woo KS, Fung KP.

Department of Biochemistry and Institute of Chinese Medicine, The Chinese University of Hong Kong, Hong Kong SAR, PR China.

The roots of Pueraria thomsonii and Pueraria lobata are officially recorded in Chinese Pharmacopoeia under the same name Radix Puerariae. However, the aqueous root extract of Pueraria lobata showed more potent antioxidant activity than that of Pueraria thomsonii. A qualitative HPLC method was developed to compare the chemical profiles of Pueraria thomsonii and Pueraria lobata, which revealed four major common peaks (daidzein 1, daidzin 2, puerarin 3 and 5-hydroxypuerarin 4) and two major different peaks (3-hydroxypuerarin 5 and 3'-methoxypuerarin 6) in their chromatograms. Semi-quantitative analysis showed that the contents of 1-3 in Pueraria lobata are about three, three, and five times higher than those of Pueraria thomsonii, respectively. The higher contents of isoflavonoids in Pueraria lobata were inferred to be responsible for its more potent antioxidant activity as compared with that of Pueraria thomsonii. The HPLC method developed in this study and chemical markers 1-6 can be used for the rapid identification and evaluation of Radix Puerariae herbs and their aqueous supplements, and the results of this investigation support the use of Pueraria lobata and Pueraria thomsonii in the clinic application and as dietary supplement, respectively.

Publication Types: PMID: 15588661 [PubMed - indexed for MEDLINE]

182. Purification of the isoflavonoid puerarin by adsorption chromatography on cross-linked 12% agarose.

He X, Tan T, Janson JC.

Department of Biotechnology, School of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.

The isoflavonoid puerarin in extracts of the well-known traditional Chinese drug Radix puerariae (root of the plant Pueraria lobata) can be separated from other isoflavonoids by adsorption chromatography on the cross-linked 12% agarose gel Superose 12 equilibrated in distilled water. The adsorption is totally quenched by the addition of 50% acetic acid. The separation of the isoflavonoids is tentatively ascribed to interaction with the residues of the cross-linking reagents used in the manufacturing process of Superose 12. Thus, no useful separation can be achieved with non-cross-linked 12% agarose gel media. Symmetric elution profiles at high sample loadings (16 mg on a 24 ml column) suggest linear adsorption isotherms for the isoflavonoids.

Publication Types: PMID: 15584227 [PubMed - indexed for MEDLINE]

183. Inhibition of Trypanosoma cruzi by plant extracts used in Chinese medicine.

Lirussi D, Li J, Prieto JM, Gennari M, Buschiazzo H, Ríos JL, Zaidenberg A.

Cátedra de Farmacología, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Calles 60 y 120, La Plata 1900, Argentina.

In this work, we assessed the effect of extracts obtained from 17 plants used in traditional Chinese medicine. These extracts were tested in vitro with the epimastigote form of Trypanosoma cruzi, clone Bra C(15) C(2), at 27 degrees C in F-29 medium at a concentration of 100 microg/ml in axenic cultures. Allopurinol was used as reference drug. Seven plant extracts showed inhibitory activities lower than 25%. Pueraria lobata, Mahonia beaei, Dictamus dasycarpus, Kochia scoparia, Sophora flavescens and Ligustrum lucidum showed effects with inhibition values between 25% and 60%, whereas Lithospermum erythrorhizon, Saussurea lappa, Melia toosendan and Cinnamomum cassia showed the greatest inhibitory activity of 100%. The IC(50) of these extracts were: 0.4, 2.4, 1.8 and 3.9 microg/ml, respectively. The MTT assay was made and did not show cytotoxic activity. These results allowed us to suggest that L. erythrorhizon, S. lappa, M. toosendan and C. cassia could be a source of new compounds against T. cruzi.

Publication Types: PMID: 15567249 [PubMed - indexed for MEDLINE]

184. [The effects of Radix Puerariae decoction on levels of antibody and interleukin-2 in mice]

[Article in Chinese]

Ma J, Cheng G, Li D, Liu H.


Dalian Medical University, Dalian 116027.

OBJECTIVE: To study the effects of Radix Puerariae on immune response. METHOD: Decoction of Radix Puerariae was administrated to mice (i.g.), and then sheep red blood cell (SRBC) and ovalbumin were used as antigens. Levels of anti-SRBC, anti-ovalbumin antibodies and serum interleukin-2 (IL-2) were determined. RESULT: Levels of anti-SRBC, anti-ovalbumin antibodies were higher in experimental group than that in control group (P < 0.05), while concertrations of IL-2 showed no significant difference. CONCLUSION: Decoction of Radix Puerariae can promote the formation of antibody.

Publication Types: PMID: 15562724 [PubMed - indexed for MEDLINE]

185. [Effects of puerarin on number and activity of endothelial progenitor cells from peripheral blood]

[Article in Chinese]

Zhang FR, Chen JZ, Zhu JH, Wang XX, Zhu JH, Shang YP, Guo XG, Dai HM, Sun J.

Department of Cardiovascular Diseases, First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310003, China.

OBJECTIVE: To investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferation, migration and adhesion. METHOD: Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days culture, attached cells were stimulated with puerarin (to make a series of final concentrations: 0. 1, 0.5, 1, 3 mmol x L(-1)) or vehicle control for the respective time points (6, 12, 24, 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, then adherent cells were counted. RESULT: Incubation of isolated human MNCs with puerarin dose increased the number of EPCs, maximum at 3 mmol x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, puerarin also promoted EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity. CONCLUSION: Puerarin can augment the number of EPCs with enhanced functional activity.

Publication Types: PMID: 15506293 [PubMed - indexed for MEDLINE]

186. [Experimental study on anti-pyretic effect of gegen qin lian decoction and its compounds]

[Article in Chinese]

Yu LZ, Wu JY, Luo JB, Huang XG, Shao HX, Lin H.

Department of Chinese Drugs Pharmacology of First Military Medical University, Guangzhou 510515, China. yulzh@fimmu.com

OBJECTIVE: To investigate composition principles of Gegen Qin Lian decoction through anti-pyretic experiment. METHOD: Pharmacological effects of different compounds of Gegen Qin Lian decoction according to six hours temperature response index (TRI6) and average top temperature response height (deltaT) after the decoction was given to feverish animal model by inactived bacteria suspension. RESULT: As for reducing six hour temperature response index, Scutellaria baicalensis root was the main effective drug. Pueraria lobata root could enforce the effect while Coptis chinensis rhizome and Glycyrrhiza uralensis root counteracted it. As for reducing average top temperature response height, the Effects of four herbal drugs were the same as for TRI6. CONCLUSION: Of the compounds of Gegen Qin Lian decoction, as to the pharmcological anti-pyretic effects, the best one is the compound of Scutellaria baicalensis and Pueraria lobata roots.

Publication Types: PMID: 15503775 [PubMed - indexed for MEDLINE]

187.Establishment of in vitro test system for the evaluation of the estrogenic activities of natural products.

Kim OS, Choi JH, Soung YH, Lee SH, Lee JH, Ha JM, Ha BJ, Heo MS, Lee SH.


Department of Bioscience and Biotechnology, Silla University, Busan 617-736, Korea.

In order to evaluate estrogenic compounds in natural products, an in vitro detection system was established. For this system, the human breast cancer cell line MCF7 was stably transfected using an estrogen responsive chloramphenicol acetyltransferase (CAT) reporter plasmid yielding MCF7/pDsCAT-ERE119-Ad2MLP cells. To test the estrogenic responsiveness of this in vitro assay system, MCF7/pDsCAT-ERE119-Ad2MLP cells were treated with various concentrations of 17beta-estradiol. Treatments of 10(-8) to 10(-12) M 17beta-estradiol revealed significant concentration dependent estrogenic activities compared with ethanol. We used in vitro assay system to detect estrogenic effects in Puerariae radix and Ginseng radix Rubra extracts. Treatment of 500 and 50 microg/ml of Puerariae radix extracts increased the transcriptional activity approximately 4- and 1.5-fold, respectively, compared with the ethanol treatment. Treatment of 500, 50, and 5 microg/ml of Ginseng radix Rubra extracts increased the transcriptional activity approximately 3.2-, 2.7-, and 1.4-fold, respectively, compared with the ethanol treatment. These observations suggest that Puerariae radix and Ginseng radix Rubra extracts have effective estrogenic actions and that they could be developed as estrogenic supplements.

Publication Types: PMID: 15473658 [PubMed - indexed for MEDLINE]

188. Antibacterial effect of crude water-soluble arrowroot (Puerariae radix) tea extracts on food-borne pathogens in liquid medium.

Kim S, Fung DY.

Department of Animal Sciences and Industry, Food Microbiology Laboratory, Kansas State University, Manhattan, KS 47907-2009, USA. kim98@purdue.edu

AIMS: To evaluate the effect of crude water-soluble arrowroot tea extracts on microbial growth of food-borne pathogens in liquid medium and to confirm the damage to bacterial cells using Transmission Electronic Microscopy (TEM). METHODS AND RESULTS: Inhibition of growth of Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Listeria monocytogenes and Staphylococcus aureus was investigated using Brain Heart Infusion (BHI) broth containing 0 (control), 0.63, 1.25, 2.5 and 5.0% (w/v) arrowroot tea. Bacterial cell counts were performed on specific selective agar on days 0, 1, 3 and 5. BHI containing 5.0% arrowroot tea extract showed a 6-7 log suppression of growth for all test strains on days 3 and 5, compared with the control. Even 0.63% arrowroot tea effectively inhibited microbial growth of all test strains on day 5. TEM images of the samples treated with 5.0% arrowroot tea revealed the rupture of cell walls and nonhomogeneous disposition of cytoplasmic materials within treated bacteria. CONCLUSIONS: Crude water-soluble arrowroot tea extract strongly inhibited microbial growth of all test pathogens in liquid medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Water-soluble arrowroot tea extract has the potential to be used directly on foods or as a spray on the surfaces of food handling and processing facilities in order to prevent microbial growth of both Gram-negative and Gram-positive bacteria.

Publication Types: PMID: 15355532 [PubMed - indexed for MEDLINE]

189. Supplementation of Pueraria radix water extract on changes of antioxidant enzymes and lipid profile in ethanol-treated rats.

Lee JS.

Department of Food and Nutrition, Kosin University, Busan 606-701, South Korea. jslee@Kosin.ac.kr

BACKGROUND: Water extract of Pueraria radix (PRWE), traditional oriental medicinal plant, may have an effect on the activity of hepatic antioxidant enzymes and lipid profile in ethanol-treated rats. METHODS: Male Sprague-Dawley rats were divided into control, ethanol, PRWE and ethanol-PRWE supplemented groups. Twenty-five percent (v/v) ethanol (5 g/kg body weight) was orally administered once a day for 5 weeks. The PRWE was supplemented in a diet based on 1500 mg of raw PRWE/kg body weight/day. RESULTS: Ethanol feeding resulted in a higher alcohol dehydrogenase (ADH) activity and lower aldehyde dehydrogenase (ALDH) activity. After PRWE supplementation, both activities were increased. The PRWE supplementation resulted in a significant decrease in the plasma and liver total cholesterol concentrations in the ethanol-treated rats. Ethanol administration significantly lowered the activities of hepatic superoxide dismutase (SOD) and catalase (CAT), whereas it increased the plasma and hepatic thiobarbituric acid reactive substances (TBARS) and the hepatic glutathione peroxidase (GSH-Px) activities. However, PRWE supplementation resulted in a significant increase in the SOD and CAT activities and a significant decrease in the TBARS and the GSH-Px activities in the ethanol-treated rats. CONCLUSION: PRWE can contribute to alleviating the adverse effect of ethanol ingestion by enhancing the lipid metabolism as well as the hepatic antioxidant defense system.

Publication Types: PMID: 15313149 [PubMed - indexed for MEDLINE]

190. Effects of puerarin on learning-memory and amino acid transmitters of brain in ovariectomized mice.

Xu X, Hu Y, Ruan Q.

College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, P.R. China. sky88@mail.zjnu.net.cn

In the present study, the protective effects of puerarin (Pur) on learning and memory in ovariectomized mice were investigated.One week after ovariectomy (Ovx) or sham operation, female mice were given a 4-week treatment of Pur (50 or 100 mg/kg,i.p.) or estradiol benzoate (EB, I or 5 pg/day, i.p.). The results showed that, following treatment with 50 or 100 mg/kg Pur in Ovx mice, the training times for achieved learning criterion in aY-maze declined by 11.8 % and 17.8 % (P > 0.05 and P < 0.05), and that memory retention increased by 23.3 % and 28.3 % (P < 0.05 and P < 0.05), respectively. In addition, the prolonged escape latency of platform finding in a water maze was shortened by 15.5% and 23.8% (P < 0.05 and P < 0.05). After a behavior test,the levels of glutamate (Glu) and GABA in the frontal cortex and hippocampus were assessed using high-performance liquid chromatography (HPLC). The results showed that 100 mg/kg Pur normalized the levels of Glu and GABA in the hippocampus of Ovx mice, while the Glu level was significantly elevated from 24.1+/- 4.5 to 27.9+/- 3.5 mg/g protein (P < 0.05), and GABA was decreased from 5.64+/- 1.06 to 4.73+/- 0.85 mg/g protein (P < 0.05).The results indicated that Pur possessed phytoestrogen activity,and long-term treatment of Pur ameliorated learning and memory deficits of Ovx mice through affecting the activity of the glutamatergic/GABAergic system in the hippocampus.

Publication Types: PMID: 15303253 [PubMed - indexed for MEDLINE]

191. Kudzu root extract suppresses voluntary alcohol intake and alcohol withdrawal symptoms in P rats receiving free access to water and alcohol.

Benlhabib E, Baker JI, Keyler DE, Singh AK.

Department of Veterinary Diagnostic Medicine, College of Veterinary Medicine, University of Minnesota, 1333 Gortner Avenue, St. Paul, MN 55108, USA.

Alcohol-preferring (P) rats, given free choice to drink water or 15% alcohol, drank 7-10 g of alcohol/kg/day, giving blood alcohol values ranging from 16 to 24 mg/dL. Body weight and food and total fluid intake values in control and alcohol-drinking P rats did not differ significantly, while water intake was inversely related to the amount of alcohol consumed. Alcohol withdrawal after 50 days of alcohol drinking caused withdrawal symptoms such as hypersensitivity, poor landing coordination, and tremors. A daily 0.5, 0.75, and 1.0 g/kg dose of kudzu root (KdR) did not affect body weight and food and water intake values in control (no alcohol) P rats. Subchronic feeding of relatively higher KdR doses (0.75 and 1.0 g/kg) caused a 25-30% reduction in weight gain. The 0.5 g/kg KdR dose caused a 50-60% reduction in alcohol consumption, abolished the development of alcohol withdrawal symptoms, but did not affect blood alcohol levels. The higher KdR doses did not further reduce alcohol consumption. Alcohol suppressed the weight-reducing effects of KdR. The KdR extract used in this study contained 150 mg/g of puerarin, 13 mg/g of daidzin, 4 mg/g of daidzein, 3 mg/g of genistin, 0.2 mg/g of genistein, and 1 mg/g of glycetin. Blood and liver samples contained mostly puerarin and a trace amount of daidzein that may have been formed by the hydrolysis of daidzin by liver enzymes. An important observation was that brain samples obtained from KdR-fed or alcohol + KdR-fed rats did not contain any of the KdR isoflavones. Thus, KdR isoflavones suppressed alcohol drinking and withdrawal symptoms without entering the brain.

PMID: 15298764 [PubMed - indexed for MEDLINE]

192. Quantitative analysis of phytoestrogens in kudzu-root, soy and spiked serum samples by high-pressure liquid chromatography.

Benlhabib E, Baker JI, Keyler DE, Singh AK.

Department of Veterinary Diagnostic Medicine, College of Veterinary Medicine, University of Minnesota, St Paul Campus, 1333 Gortner Ave, St Paul, MN 55108, USA.

A sensitive and reliable HPLC method that allows simultaneous quantification of phytoestrogens extracted from kudzu-root and soy preparations, and serum samples has been developed. Kudzu-root and soy preparations were mixed with 5 microg flavone and 15 microg rutin (internal standards) and the phytoestrogens were extracted by using solid-phase (C18) extraction cartridges. Blank or spiked serum samples were extracted by using either C18 cartridges or trichloroacetic acid-methanol extraction. The extracts were analyzed by the HPLC equipped with a reverse-phase (250 x 4 mm, C18) column and UV, diode-array or MS detector. A linear gradient of acetic acid and acetonitrile provided excellent separation of glycoside and aglycone-phytoestrogens from kudzu root and soy preparations. The C18 cartridge extraction of serum yielded excellent recovery of both glycoside- and aglycone-phytoestrogens, while the trichloroacetic acid-methanol extraction yielded excellent recovery of glycoside but poor recovery of aglycone compounds. UV and MS detectors were suitable for phytoestrogen analysis in plant and serum samples, while the diode-array detector was suitable for generating the UV absorbance curve for phytoestrogens.

Publication Types: PMID: 15273977 [PubMed - indexed for MEDLINE]

193. Anti-inflammatory and anti-nociceptive effects of the extract from Kalopanax pictus, Pueraria thunbergiana and Rhus verniciflua.

Kim IT, Park YM, Shin KM, Ha J, Choi J, Jung HJ, Park HJ, Lee KT.

Department of Biochemistry, College of Pharmacy, Kyung-Hee University, Seoul 130-701, Republic of Korea.

The combined extracts obtained from three Chinese herb medicine, Kalopanax pictus, Pueraria thunbergiana and Rhus verniciflua, have been used as therapeutics for diabetes mellitus in Korea. In the present study, we have investigated their possible anti-inflammatory effects by comparing the potency of individual extracts with that of the combined extracts. An individual water extract prepared from Kalopanax pictus, Pueraria thunbergiana, and Rhus verniciflua was named K-1, P-1, and R-1, respectively. Simultaneously, we also prepared the combined extracts from above three plant materials by identical methods and named KPR-1. These four extracts were further fractionated into the EtOAc extracts, and these were designated as K-2, P-2, R-2, and KPR-2, respectively. These eight samples were subjected to the nitrite assays in LPS-induced macrophage 264.7 cells. KPR-2 exhibited the most pronounced effect on the inhibition of NO production among all the extracts. KPR-2 also significantly decreased PGE2, and TNF-alpha release. In addition, KPR-2 showed in vivo anti-inflammatory activity against acute paw edema induced by carrageenan in rats. When analgesic activity was measured by the acetic acid-induced abdominal constriction and hot plate test, KPR-2 showed a dose-dependent inhibition in animal models. These results suggested that the mixture extract and successive fractionation could lead to the better use of anti-inflammatory medicinal crude drugs.

Publication Types: PMID: 15261979 [PubMed - indexed for MEDLINE]

194. Passive cutaneous anaphylaxis-inhibitory action of tectorigenin, a metabolite of tectoridin by intestinal microflora.

Park EK, Shin YW, Lee HU, Lee CS, Kim DH.

Department of Biochemistry, Kon Kuk University, Chungju, Chungcheongbuk-do, Korea.

Tectoridin isolated from the flowers of Pueraria thunbergiana (Leguminosae) are metabolized to tectorigenin by human intestinal microflora. When tectoridin was orally administered to rats, tectorigenin, but not tectoridin, was detected in urine after beta-glucuronidase hydrolysis. The main metabolite tectorigenin potently inhibited the passive cutaneous anaphylaxis reaction and inhibited in vitro the release of beta-hexosaminidase from RBL-2H3 cells induced by IgE. These results suggest that tectoridin is a prodrug, which can be transformed into the active agent tectorigenin by human intestinal bacteria and can be a candidate for antiallergic agent.

PMID: 15256747 [PubMed - indexed for MEDLINE]

195. Activation of alpha1A-adrenoceptors by genistein at concentrations lower than that to inhibit tyrosine kinase in cultured C2C12 cells.

Jou SB, Huang CC, Liu IM, Cheng JT.

Department of Neurology, School of Medicine and Hospital, China Medical University, Taichung City, Taiwan, R.O.C.

Genistein, an isoflavonoid natural product, is widely used to inhibit protein tyrosine kinase (PTK). In the present study, we investigated the possible influence of genistein on alpha (1)-adrenoceptors (AR) in cultured C2C12 cells. Genistein enhanced the uptake of radioactive glucose into C2C12 cells in a concentration-dependent manner. Similar results were also observed in samples treated with daidzein, the inactive congener for PTK inhibition. The effect of genistein on alpha (1)-AR was further characterized using the displacement of [ (3)H]prazosin binding in C2C12 cells. The increase in radioactive glucose uptake by genistein was abolished by RS17053 at a concentration sufficient to block alpha (1A)-AR. The pharmacological inhibition of phospholipase C (PLC) by U73122 resulted in a concentration-dependent reduction of genistein-stimulated glucose uptake in C2C12 cells. This inhibition by U73122 was specific because the inactive congener, U73343, failed to modify the action of genistein. Moreover, genistein can activate alpha (1A)-AR at a concentration (1 micromol/L) lower than that (50 micromol/L) needed to abolish the insulin-stimulated phosphorylation of PTK. The obtained data indicate an activation of alpha (1A)-AR by genistein to increase the glucose uptake into C2C12 cells and this supports the application of genistein as a TK inhibitor.

Publication Types: MID: 15254853 [PubMed - indexed for MEDLINE]

196. The differential anti-proliferation effect of white (Pueraria mirifica), red (Butea superba), and black (Mucuna collettii) Kwao Krua plants on the growth of MCF-7 cells.

Cherdshewasart W, Cheewasopit W, Picha P.

Department of Biology, Faculty of Science, Chulalongkorn University, Phyathai Road, Bangkok 10330, Thailand. cwichai@sc.chula.ac.th

The differential anti-proliferation effect of white (Pueraria mirifica), red (Butea superba) and black (Mucuna collettii) Kwao Krua plant extracts on the growth of MCF-7 cells was evaluated after 4 days of incubation. The percent cell growth comparison was based on protein determination of the harvested cells in parallel with the control group and Pueraria lobata treatment group. Pueraria lobata led to no proliferation and a mild anti-proliferation effect on the growth of MCF-7 cells. Pueraria mirifica caused proliferation at 1 microg/mL and an anti-proliferative effect on the growth of MCF-7 cells at 100 and 1000 microg/mL with an ED50 value of 642.83 microg/mL. Butea superba led to no proliferation and an anti-proliferation effect on the growth of MCF-7 cells at 10, 100 and 1000 microg/mL with an ED50 value of 370.91 microg/mL. Mucuna collettii led to no proliferation and an anti-proliferation effect on the growth of MCF-7 cells at 100 and 1000 microg/mL with an ED50 value of 85.36 microg/mL. The results demonstrated that only Pueraria mirifica showed an estrogenic effect on MCF-7 cell growth and a clear antagonistic effect with E2 at high concentration. Butea superba and Mucuna collettii exhibited only anti-proliferation effects on the growth of MCF-7 cells in relation with a possible anti-estrogen mechanism or a potent cytotoxic effect.

Publication Types: PMID: 15234761 [PubMed - indexed for MEDLINE]

197. Effects of Radix Puerariae flavones on liver lipid metabolism in ovariectomized rats.

Wang JF, Guo YX, Niu JZ, Liu J, Wang LQ, Li PH.

Department of Biochemistry, Beijing University of Chinese Medicine, Beijing 100029, China.

AIM: To study the effects of Radix Puerariae flavones (RPF) on liver lipid metabolism in ovariectomized (OVX) rats. METHODS: Forty adult female Wistar rats were randomly divided into four groups: OVX group; sham-OVX group; OVX+estrogen group and OVX+RPF group. One week after operation rats of the first two groups were treated with physiological saline, rats of OVX+estrogen group with estrogen (1 mg/kg.b.w.) and rats of OVX+RPF group with RPF (100 mg/kg.b.w.), respectively for 5 weeks. After the rats were killed, their body weight, the weight of the abdominal fat and uterus were measured, and the levels of total cholesterol (TC) and triglyceride (TG) in liver homogenate were determined. RESULTS: Compared with the sham-OVX group, the body mass of the rats in OVX group was found increased significantly; more abdominal fat in store; TC and TG in liver increased and uterine became further atrophy. As a result, the RPF was found to have an inhibitive action on those changes of various degrees. CONCLUSION: RPF has estrogen-like effect on lipid metabolism in liver and adipose tissue.

PMID: 15222048 [PubMed - indexed for MEDLINE]

198. The antidepressant effect of ethanol extract of radix puerariae in mice exposed to cerebral ischemia reperfusion.

Yan B, Wang DY, Xing DM, Ding Y, Wang RF, Lei F, Du LJ.


Institute of Medicinal Plant, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100094, China.

In our pilot study, the depressive-like behaviors of mice exposed to cerebral ischemia reperfusion (CIR) were observed and the antidepressant effects of radix puerariae (RP; root of the Pueraria plant) extract in CIR mice were assessed because it was speculated that the neuronal damage caused by CIR played an important role in the development of poststroke depression (a common and severe complication after stroke) and the RP extract was reported to exhibit effect of neuronal protection from cerebral ischemia damage. Our studies above indicated that the RP extract markedly shortened the increased immobility time induced by CIR of male mice in the forced swimming test (FST) and tail suspension test (TST), indicating a possible antidepressant activity. Thus, the aim of the present study was to confirm the putative antidepressant effect of RP extract (75, 150, and 300 mg/kg, administered orally 24 h after the CIR) on reserpine-induced symptoms. To get further insight into the mode of antidepressant action of RP extract, biochemical examination was conducted concomitantly to examine possible involvement of the brain monoamine systems in the behavioral syndromes observed. In CIR mice, pronounced low levels of norepinephrine (NE) and 4-dihydroxyphenylacetic acid (DOPAC, a metabolite of dopamine) in the hippocampus or striatum were detected, which were reversed by RP extract, whereas no significant change of serotonin (5-HT) was detected in either CIR or RP extract-treated mice. The data suggested that the disturbance of NE and DA systems in hippocampus and striatum played more important roles in the development of depressive-like behavior of CIR mice than 5-HT system did, and RP extract ameliorated the abnormal symptoms caused by CIR, which may throw new lights on the treatment of poststroke depression.

Publication Types: PMID: 15219773 [PubMed - indexed for MEDLINE]

199. Scale-up production of puerarin from hairy roots of Pueraria phaseoloides in an airlift bioreactor.

Kintzios S, Makri O, Pistola E, Matakiadis T, Shi HP, Economou A.

Laboratory of Plant Physiology, Faculty of Agricultural Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece. skin@aua.gr

Hairy roots of the Chinese herb, Pueraria phaseoloides, obtained from leaf explants and transformed with the Agrobacterium rhizogenes, were cultured in 2.5 l airlift bioreactors for three weeks. Puerarin accumulated at 5,570 microg g(-1) dry wt, which is near 200 times as much as in 250 ml flask cultures. In addition, puearin was exuded into the nutrient medium at final concentrations higher than in the hairy roots themselves.

Publication Types: PMID: 15218379 [PubMed - indexed for MEDLINE]

200. Identification of puerarin and its metabolites in rats by liquid chromatography-tandem mass spectrometry.

Prasain JK, Jones K, Brissie N, Moore R, Wyss JM, Barnes S.

Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA. jprasain@uab.edu

Puerarin (daidzein-8-C-glucoside) is the major bioactive isoflavone of kudzu root (the root of Pueraria lobata). Its metabolic fate, however, is not well-known. In this study, a sensitive and specific LC-ESI-MS/MS method for the determination of puerarin and its metabolites daidzein, dihydrodaidzein, and equol was developed for their analysis in biological samples. Two new metabolites of puerarin, mono- and dihydroxylated derivatives, were detected in the urine and feces of rats after oral administration. The persistence of puerarin in blood and urine as the principal metabolic form for the period of 4-72 h after oral administration suggested that puerarin is rapidly absorbed from the intestine without metabolism. Its presence in organs such as the brain suggests that this glucoside may enter tissues by specific transport pathways. Study of these metabolites may provide further understanding of the health beneficial effects of puerarin in kudzu dietary supplements.

Publication Types: PMID: 15186086 [PubMed - indexed for MEDLINE]


 

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