Constituent isoflavones of Puerariae radix as a potential neuroprotector in cognitive impairment: Evidence from preclinical studies.

With the increasing aging population worldwide, the incidence of senile cognitive impairment (CI) is increasing, posing a serious threat to the health of elderly persons. Despite developing new drugs aimed at improving CI, progress in this regard has been insufficient. Natural preparations derived from plants have become an unparalleled resource for developing new drugs. Puerariae radix (PR) has a long history as Chinese herbal medicine. PR is rich in various chemical components such as isoflavones, triterpenes, and saponins. The isoflavones (puerarin, daidzein, formononetin, and genistein) exhibit potential therapeutic effects on CI through multiple mechanisms. Relevant literature was organized from major scientific databases such as PubMed, Elsevier, SpringerLink, ScienceDirect, and Web of Science. Using "Puerariae radix," "Pueraria lobata," "isoflavones," "puerarin," "antioxidant," "daidzein," "formononetin," "genistein," "Alzheimer"s disease," and "vascular cognitive impairment" as keywords, the relevant literature was extracted from the databases mentioned above. We found that isoflavones from PR have neuroprotective effects on multiple models of CI via multiple targets and mechanisms. These isoflavones prevent Aß aggregation, inhibit tau hyperphosphorylation, increase cholinergic neurotransmitter levels, reduce neuroinflammation and oxidative stress, improve synaptic plasticity, promote nerve regeneration, and prevent apoptosis. PR has been used as traditional Chinese herbal medicine for a long time, and its constituent isoflavones exert significant therapeutic effects on CI through various neuroprotective mechanisms. This review will contribute to the future development of isoflavones present in PR as novel drug candidates for the clinical treatment of CI.

Subacute oral toxicity and bacterial mutagenicity of a mixture of Puerariae radix and Hizikia fusiforme extracts.

The study aims to identify the safety profile of a mixed extract (KGC-02-PS) from two traditional medicinal herbs, Puerariae radix and Hizikia fusiforme. In a subacute oral toxicity study, KGC-02-PS was administered orally for 28 days by gavage to Sprague Dawley rats (both sexes) at a daily dose of 0, 500, 1000, and 2000 mg/kg body weight. Bodyweight, food consumption, and clinical signs were monitored during the experimental period. After administering the final dose, this study conducted hematology, serum biochemistry, and pathological evaluations. In addition, the study performed a bacterial reverse mutation test with varying concentrations of KGC-02-PS (312.5 µg - 5,000 µg/plate) following OECD guideline No. 471, before testing five bacterial strains (Salmonella typhimurium TA98, TA100, TA1535, TA1537, and Escherichia coli WP2) in the presence or absence of metabolic activation. The preclinical evaluation of KGC-02-PS's subacute oral toxicity yielded no associated toxicological effects or any changes in clinical signs, body weight, and food consumption. Moreover, examining KGC-02-PS's hematological and serum biochemical characteristics and pathology yielded no toxicological changes in terms of organ weight measurements and gross or histopathological findings. KGC-02-PS neither increased the number of revertant colonies in all bacterial strains used in the bacterial reverse mutation test, nor did it induce genotoxicity related to bacterial reverse mutations under the study's conditions. Also, KGC-02-PS's no-observed-adverse-effect level was greater than 2000 mg/kg.

Chronoeffects of the Herbal Medicines Puerariae radix and Coptidis rhizoma in Mice: A Potential Role of REV-ERBα.

Identifying drugs with dosing time-dependent effects (chronoeffects) and understanding the underlying mechanisms would help to improve drug treatment outcome. Here, we aimed to determine chronoeffects of the herbal medicines Puerariae radix (PR) and Coptidis rhizoma (CR), and investigate a potential role of REV-ERBα as a drug target in generating chronoeffects. The pharmacological effect of PR on hyperhomocysteinemia in mice was evaluated by measuring total homocysteine, triglyceride levels and lipid accumulation. PR dosed at ZT10 generated a stronger effect on hyperhomocysteinemia than drug dosed at ZT2. Furthermore, PR increased the expression levels of REV-ERBα target genes Bhmt, Cbs and Cth (encoding three key enzymes responsible for homocysteine catabolism), thereby alleviating hyperhomocysteinemia in mice. Moreover, CR attenuated chronic colitis in mice in a dosing time-dependent manner based on measurements of disease activity index, colon length, malondialdehyde/myeloperoxidase activities and IL-1ß/IL-6 levels. ZT10 dosing generated a stronger anti-colitis effect as compared to ZT2 dosing. This was accompanied by lower production of colonic inflammatory cytokines (i.e., Nlrp3, IL-1ß, IL-6, Tnf-α and Ccl2, REV-ERBα target genes) in colitis mice dosed at ZT10. The diurnal patterns of PR and CR effects were respectively consistent with those of puerarin (a main active constituent of PR, a REV-ERBα antagonist) and berberine (a main active constituent of CR, a REV-ERBα agonist). In addition, loss of Rev-erbα in mice abolished the dosing time-dependency in PR and CR effects. In conclusion, the therapeutic effects of PR and CR depend on dosing time in mice, which are probably attributed to diurnal expression of REV-ERBα as the drug target. Our findings have implications for improving therapeutic outcomes of herbal medicines with a chronotherapeutic approach.

Comprehensive study of the interaction between Puerariae Radix flavonoids and DNA: From theoretical simulation to structural analysis to functional analysis.

Puerariae Radix (PR) is a natural herb whose active ingredient is mainly flavonoids. To explore the interaction between PR flavonoids and DNA not only has important biological implications for understanding the mechanism of action, but also helps develop PR products for the design of appropriate dietary interventions to aid cancer treatment. In this work, we comprehensively studied the interaction between six kinds of PR flavonoids and DNA from four different and progressive levels, including molecular docking, multi-spectral analysis, and functional analysis in vitro and in cell. Results show that the DNA binding affinity of six flavonoids is in an order of quercetin > formononetin > daidzein > puerarin > 4'-methoxy puerarin > puerarin 6″-O-xyloside (POS), in which quercetin can significantly inhibit DNA amplification owing to its strongest binding affinity. The binding between quercetin and DNA is further revealed to be intercalated binding, which can cause conformational changes in DNA, thereby exhibiting an activity of cell cycle arrest and anti-proliferative. This property of quercetin can be utilized for the further development of flavonoids with anticancer activity. In addition to the potential application, this work also provides a platform for the comprehensive study of the interaction between micromolecules and DNA.

Puerariae Radix Prevents Anxiety and Cognitive Deficits in Mice Under Oligomeric Aß-Induced Stress.

To evaluate the therapeutic effects of Chinese herbal medicine (CHM) for Alzheimer's disease (AD), we evaluated five CHMs in oligomeric Aß25-35-treated mouse primary hippocampal neuronal cultures. The aqueous extract from the root of Pueraria lobata (Puerariae Radix; PR) showed better neuroprotective effects than did the other four CHM aqueous extracts, including Gardenia jasminoides, Eleutherococcus senticosus, Rhodiola rosea, and Panax, in the primary culture treated with saline or oligomeric Aß25-35. Furthermore, the neuroprotective effects of aqueous extract of PR were also better than its well-known active compound, puerarin, against the neurotoxicity of oligomeric Aß25-35 in a primary culture. For in vivo experiments, C57BL/6J male mice that received direct infusion of soluble oligomeric Aß25-35 into the bilateral hippocampal CA1 subregion were used as an alternative AD mouse model. The effects and molecular mechanisms of chronic systemic administration of PR aqueous extract were evaluated in the alternative AD model. PR aqueous extract prevented anxiety and cognitive impairment in mice associated with a decrease in the levels of Aß deposition, tau protein phosphorylation, inflammation, loss of noradrenergic, and serotonergic neurons and an increase in the levels of synaptophysin and insulin degrading enzyme (IDE) against the toxicity of oligomeric Aß25-35. Furthermore, no obvious damage to the liver and kidney was detected after chronic systemic administration of PR aqueous extract. Therefore, using PR could be a safer, more effective therapeutic strategy than using its active compound puerarin to prevent both cognitive and noncognitive dysfunction and related pathological features of AD.

Screening and identification of the active components from Puerariae Radix by HUVEC/CMC-LC-MS2.

Puerariae Radix (PR) serves as food and medicinal plant for thousands of years with explicit efficacy for heart diseases, while biological target specifically binding-oriented screening of the active components in PR remains a preliminary stage. Cell membrane chromatography (CMC) is newly developed approach where interactions between active components and certain biological targets can be effectively studied, Human umbilical vein endothelial cell (HUVEC) membrane, with its abundant receptors such as ß and AT1, is most eligible for constructing CMC. In this study, an HUVEC/CMC-LC-MS2 system was developed for screening active components in PR, 11 compounds were screened out and four of them were identified. Besides puerarin, the rest identified are daidzin, pueroside D and 3'-hydroxypuerarin. The study provides more reference for CMC applications and PR exploitation.

Enhancement of the antioxidant and hypolipidemic activities of Puerariae radix by fermentation with Aspergillus niger.

Puerariae radix (PR) is a traditional Chinese food and medicine. In this study, the chemical profile and bioactivities of PR fermented with Aspergillus niger (PFA) were investigated. Based on HPLC analysis, PFA chemical profile changed and total phenols increased by 12.5% relative to PR. Consequently, the in vitro antioxidant activity significantly improved. According to the blood lipid analysis in mice, PFA showed a better ability to inhibit the increased blood lipid levels induced by Triton WR-1339 than PR. In a quantitative real-time PCR (qRT-PCR) study, PFA up-regulated cholesterol 7-alpha hydroxylase (CYP7A1) and low-density lipoprotein receptor mRNA expression, which were 50% and 44.8% higher than the levels induced by high-dose PR. Conversely, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCoAR) mRNA expression was down-regulated to 46.8% lower than the levels induced by high-dose PR. The qRT-PCR results suggested that PFA displayed better hypolipidemic activity than PR, due to its superior ability to regulate mRNA expression.

Comparative pharmacokinetics of major bioactive components from Puerariae Radix-Gastrodiae Rhizome extracts and their intestinal absorption in rats.

Puerariae Radix (PR) and Gastrodiae Rhizome (GR) is frequently used in traditional herbal formulas to treat cardio-cerebral vascular diseases due to their synergistic effects. In this study, to elucidate the action mechanism of PR-GR in vivo, a simple and reliable ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of nine bioactive ingredients from PR-GR in plasma was developed and applied to a comparative pharmacokinetic study following oral administration of PR, GR, and PR-GR aqueous extracts in rats. The effect of GR on the absorption of components of PR was also investigated by single-pass intestinal perfusion study. Results showed that comparing to the single herbs, PR-GR extract significantly increased the systemic exposure of puerarin, 3'-hydroxypuerarin, 3'-methoxypuerarin, 6″-O-xylosylpuerarin, daidzin, genistein, and gastrodin. Moreover, the intestinal absorption of puerarin and daidzin could be improved by GR extract and inhibitors of P-glycoprotein and multidrug resistanceassociated protein 2, respectively. These results indicate that the combination of PR and GR increases the levels of their bioactive ingredients exposed in the blood, and GR increases the absorption of ingredients of PR may by inhibition of the efflux mediated by P-glycoprotein and multidrug resistanceassociated protein 2. This is the first report for the pharmacokinetics and intestinal absorption of PR-GR, which may explain their synergetic effects in the treatment of circulatory systematic diseases and provide a meaningful insight for their clinical applications.

Simulation and prediction of material foundation of Puerariae Radix for ischemic stroke based on molecular docking technology / 中草药

Objective To study the mechanism of Puerariae Radix in the treatment of ischemic stroke by using molecular docking technology. Methods The small molecules of Puerariae Radix based on molecular docking technology docked with 21 key targets protein of cerebral ischemic stroke, and multi-component protein target network was established by Cytoscape 3.1.1 software. Results Through virtual screening of molecular docking technology, 28 active small molecules of Puerariae Radix were chosen, 12 of which were novel small molecules, and it identified that 11 of those compounds had strong interactions with no less than 10 targets. Conclusion The molecular docking can be used to find the active components of Puerariae Radix in treatment of cerebral ischemic stroke, which provide a new reference of studying on the multiple ingredients and targets of Chinese materia medica compounds.

Pueraria lobate Inhibits RANKL-Mediated Osteoclastogenesis Via Downregulation of CREB/PGC1ß/c-Fos/NFATc1 Signaling.

Puerariae radix, the dried root of Pueraria lobate Ohwi, is known to prevent bone loss in ovariectomized mice; however, the precise molecular mechanisms are not understood. In this study, we investigated the effects and underlying mechanisms of action of Puerariae radix extract (PRE) on receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis. PRE dose-dependently inhibited osteoclast differentiation and formation, decreased the bone-resorbing activity of osteoclasts, and downregulated the expression of osteoclast differentiation marker genes. The expression of osteoclastogenic factors produced by PRE-treated osteoblasts such as RANKL, macrophage colony-stimulating factor (M-CSF), and osteoprotegerin (OPG) was comparable to that of untreated (control) cells. However, the formation of osteoclasts via bone marrow cell and calvaria-derived osteoblast co-cultures was suppressed by PRE treatment. Therefore, the inhibitory effects of PRE on osteoclastogenesis clearly targeted osteoclasts, but not osteoblasts. PRE treatment considerably reduced RANKL-induced mitogen-activated protein kinases (MAPKs) activity, especially c-Jun N-terminal kinase, in osteoclast precursor cells. In addition, PRE markedly suppressed cAMP response element-binding protein (CREB) activation and the induction of peroxisome proliferator-activated receptor gamma coactivator 1ß (PGC1ß), which stimulate osteoclastogenesis - an effect that was not observed for puerarin and 17-ß estradiol. Finally, PRE treatment significantly repressed the expression of c-Fos and the nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), which is a master transcription factor for osteoclastogenesis in vitro and in vivo. Overall, these results strongly suggest that PRE is an effective inhibitor of RANKL-induced osteoclastogenesis and may be a potent therapeutic agent for bone-related diseases such as osteoporosis, rheumatoid arthritis, and periodontitis.

Inhibitory effect of Puerariae radix flavones on platelet-derived growth factor-BB-induced proliferation of vascular smooth muscle cells via PI3K and ERK pathways.

Abnormal proliferation of vascular smooth muscle cells (VSMCs) results in intimal thickening of the aorta, which may lead to arteriosclerosis. Therefore, VSMC antiproliferative agents may be efficient in the prevention and treatment of arteriosclerosis. Puerariae radix (PR) is the dried root of Pueraria lobata Ohwi or Pueraria thomsonii Benth. Flavones are the main components of PR and have been shown to have a protective effect on vascular disorders in traditional Chinese medicine treatments. However, the underlying molecular mechanism remains unclear. The aim of the present study was to explore the effect of PR flavone (PRF) on platelet-derived growth factor (PDGF)-BB-induced VSMC proliferation. PDGF-BB (25 ng/ml) and different doses of PRF (10, 50, 100 and 200 ng/ml) were used to treat VSMCs. The results revealed that PRF notably inhibited the PDGF-BB-induced VSMC proliferation and induced a cell cycle arrest at growth 1 phase of the cell cycle. In addition, cell cycle-associated proteins, including cyclin D1, proliferating cell nuclear antigen and cyclin-dependent kinase 4, were found to be downregulated. Furthermore, PRF inhibited the PDGF-BB-stimulated downregulation of VSMC markers, including α-smooth muscle actin, desmin and smoothelin. PDGF-BB upregulated the phosphorylation levels of phosphatidylinositide 3-kinase (PI3K) and extracellular signal-regulated kinase (ERK), which are associated with cell proliferation; however, these were decreased following PRF treatment. These observations indicated that PRF had a suppressive effect on PDGF-BB-induced VSMC proliferation by inhibiting PI3K and ERK pathways.

Rapid analysis on flavonoids in Gegen Zhiju Soft Capsule by UPLC/Q-TOF-MS / 中草药

Objective: To identify the flavonoids in Gegen Zhiju Soft Capsule (a soft capsule prepared with Puerariae Radix and Hoveniae Dulcis Fructus seu Semen and so on) using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Methods: The analysis was performed on Agilent Poroshell 120 SB-C18 column (100 mm ×2.1 mm, 2.7 μm) gradient elution with a mobile phase of 0.1% formic acid aqueous solution and acetonitrile. The flow rate was at 0.4 mL/min. TOFMS was applied for qualitative analysis, and the data were collected by the negative ion mode using ESI ion source. The parameters of ion source were as follows: drying gas temperature, 300 ℃; drying gas flow rate, 10 L/min; nebulizer gas pressure, 276 kPa, sheath gas temperature, 350 ℃; sheath gas flow rate, 11 L/min; capillary voltage, 3 500 V; fragmentor voltage, 175 V. Results: Twenty flavonoid compounds were identified by TOF-MS and literature data. Relative molecular weight was concentrated within 250-650.Conclusion: The rapid separation with UPLC and Q-TOF-MS determination of the precise molecular weight information could effectively analyze and identify the flavonoid compounds in Gegen Zhiju Soft Capsule. The method is accurate, rapid, and sensitive, and could provide a reliable and effective technique for the quality control.

Study on characteristics and dissolution behavior in decoction for boiling powders of Chinese materia medica / 中草药

Objective: To study the powder characteristics and dissolution behavior of the composition in decoction for boiling powders of Chinese materia medica, and to provide scientific evidence for particle size control and its application. Methods: Four kinds of herbal pieces such as Scutellariae Radix, Coptidis Rhizoma, Puerariae Radix, and Glycyrrhizae Preparata Radix were prepared in powders. Powder characteristics including particles distribution and mobility, and dispersion properties of particles in the water decoction were investigated. Additionally, dissolution behaviors of the composition in decoction made from powders and pieces were compared. Results: Well-distributed particles and high mobility were observed in Scutellariae Radix, Coptidis Rhizoma, and Glycyrrhizae Radix Preparata powders, rather than in Puerariae Radix powder. The suspended particles in the decoction in both powders and pieces were submicron. However, optical and electrical properties showed that the suspended particles in the decoction made from powders were more, bigger, and easier to settle as heterogeneous liquid was. Dissolution velocity and amount of the active ingredients in the decoction made from powders were significantly higher than those of pieces. Conclusion: As a new type of herbal decoction which could increase the dissolution of active ingredients, it could shorten decocting duration and improve the availability of herbs. The boiling powders are worth being studied and promoted.

Protective effects of puerariae radix extract and its single compounds on methylglyoxal-induced apoptosis in human retinal pigment epithelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: In Korea, Puerariae radix is a medicinal plant traditionally used to treat various diseases including diabetes mellitus. To provide pharmacological basis for Puerariae radix in the treatment of diabetes, we investigated the protective effects of the ethanolic extract and its single compounds on apoptosis associated with glycation in human retinal pigment epithelial (RPE) cells. MATERIALS AND METHODS: In the present work, a quantified ethanolic extract or single compounds of Puerariae radix were selected to determine its anti-apoptotic effect in human RPE cells cultured with methylglyoxal (MG), which is a stimulator of glycation. To assess the protective effect of the extract or single compounds, the cytotoxicity assessment was performed using an MTT assay in the human RPE cells. Selected active compounds or extracts were tested by FACS analysis with annexin V staining for apoptosis. RESULTS: Daidzein (1), daidzin (2), puerarin (3), 3'-hydroxy-daidzein 8-C-apiosyl (1→6) glucoside (4), and daidzein 8-C-apiosyl (1→6) glucoside (5), and pueroside B (6) were isolated from an ethanolic extract of Puerariae radix. MG-induced apoptosis was completely inhibited by Puerariae radix, ethanolic extract, and single compounds. Of the six major compounds, daidzin (2) and 3'-hydroxy-daidzein 8-C-apiosyl (1→6) glucoside (4) significantly inhibited MG-induced apoptosis. CONCLUSION: Our results provide the first evidence that, due to its anti-glycation effect, Puerariae radix extract could inhibit MG-induced apoptosis in the cultured human RPE cells. These data suggest that Puerariae radix extract, especially its single compounds daidzin and 3'-hydroxy-daidzein 8-C-apiosyl (1→6) glucoside, has potential utility as a preventive agent for glycation-related diabetic retinopathy.

Metabolic differentiations of Pueraria lobata and Pueraria thomsonii using ¹H NMR spectroscopy and multivariate statistical analysis.

Puerariae Radix was a widely used herbal medicine. Pueraria lobata (PL) and Pueraria thomsonii (PT) were the two authorized sources of Puerariae Radix (gegen) in China. In this study, metabolic differentiations between these two species were investigated using NMR spectroscopy followed by principal components analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The content of puerarin in PL and PT was also determined using quantitative (1)H NMR spectroscopy. Thirteen isoflavones were tentatively identified based on 1D and 2D NMR spectroscopic data in two species. The (1)H NMR spectra of PL and PT were obviously different. PL and PT could also be markedly discriminated from (1)H NMR spectroscopic data by PCA and PLS-DA. For the crude drug resources, isoflavones, in which puerarin is the most important one, were regarded as the reasonable markers for the discrimination of the two species. The contents of puerarin and total isoflavones in PL were quantitated much higher than those in PT. Above all, (1)H NMR spectroscopy, which can provide comprehensive profiles of the metabolites and achieve convenient determinations of puerarin and total isoflavones in a single run, is an efficient means for evaluating the medicinal samples and achieving a better quality control of Puerariae Radix.

Preparation and dissolution of Puerariae Radix Flavones Dropping Pills / 中草药

Objective: To prepare Puerariae Radix Flavones Dropping Pills and to investigate the dissolution. Methods: Exterior quality, weight variation, and resolving time were used as comprehendsive evaluation indicators to select dropping conditions by orthogonal design. HPLC was used to determine the content of puerarin and rotating basket method was used to determine the dissolution rate in vitro of the dropping pills and tablets. Results: The optimal preparation conditions for the pills were as follows: proportion of drug and matrix was 1:3, drop rate was 20 d/min, the temperature of drug fluids was 80°C, the condensate tube outlet temperature was 50°C, dimethylsilicone was the refrigerant at the temperature of 10°C and 6 cm distance above liquid level. The content of peurarin in the dropping pills was 5.542 mg/g. The accumulated dissolution rate of puerarin in the dropping pills reached 98.81% in 20 min, while the accumulated dissolution rate of puerarin in tablets was only 10.70% in 20 min. Conclusion: The preparation process and HPLC determination method are simple, stable, and feasible. The dissolution rate of puerarin can be improved in the Dropping pills.

Study on effects of puerariae radix flavones on the proliferation of multiple myeloma cell lines U266 and RPMI 8226 / 白血病·淋巴瘤

Objective To investigate the effects on proliferation of multiple myeloma cell lines U266 and RPMI 8226 induced by puerariae radix flavones (PRF) in vitro and its possible mechanism.Methods Exposed to 0,10,30,50,100 μg/ml PRF for 48 h and 72 h,the U266 and RPMI 8226 cells proliferation inhibitory rates were detected by MTT assay,cell cycles by flow cytometry (FCM),morphologic changes of U266 cells by Wright' s staining,and early-stage apoptotic rates of U266 cells by FITC-Annexin V/PI staining with FCM.Analysis of DNA fragment was made to test characteristic apoptosis DNA ladder in U266 cells.Results 0,10,30,50,100 μg/ml PRF could inhibit the proliferation of U266 and RPMI 8226 cells in a dose-dependent manner (U266 ＞ RPMI 8226).Cell cycle analyses in U266 and RPMI 8226 cells showed that sub-diploid peaks,but cell cycles changed minor.Wright's staining of U266 cells showed hardly any apoptostic character istic.Annexin V/PI double staining indicated that early-stage apoptotic rates of U266 cells exposed to 0,10,30,50,100 μg/ml PRF for 48 h were mildly increased in a dose-dependent manner.They were (3.20±0.36) ％,(5.20±0.92) ％,(7.30±1.22) ％,(8.10±0.53) ％ and (10.80±0.90) ％,respectively.The group differences had statistical significance (P ＜ 0.05).Analysis of DNA fragment barely exhibited the characteristic DNA ladder in U266 cells.Conclusion A certain concentrations of PRF could inhibit the proliferation of U266 and RPMI 8226 cells significantly.It is suggested that apoptosis related to the proliferative inhibition mechanism induced by PRF in U266 cell line,but not main.Other pathways such as necrosis and autophagy whether or not involved need further investigation.

Antioxidant and anti-fatigue activities of flavonoids from Puerariae radix.

This study evaluated the antioxidant and anti-fatigue activities of flavonoids from Puerariae radix (FPR). In vitro antioxidant activities of FPR were investigated through hydroxyl and superoxide radical scavenging activities. In vivo anti-fatigue activity of FPR was investigated through loaded swimming exercise of mice. Results showed that FPR had not only in vitro antioxidant activities, but also an in vivo anti-fatigue activity in mice. FPR possessed superoxide and hydroxyl radical scavenging activity in in vitro experimental studies. In vivo experimental studies, FPR could evidently extend exhaustive swimming time of mice, inhibit the increase of blood lactic acid (BLA), decrease serum urea nitrogen (BUN) and malondialdehyde (MDA) contents, promote increases in the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX) of mice after swimming. The results provided an important basis for developing the FPR as a novel antioxidant and anti-fatigue compound.

Effects of Supplementation of Puerariae Radix Ethanol Extract on the Antioxidative Defense System in Rats

This study was performed to investigate the effect of Puerariae radix-ethanol extracts rich in isoflavone on the antioxidative system of rats. For this purpose, first, Puerariae radix was extracted with ethanol, and its total isoflavone and puerarin contents were analysed. Second, female Sprague Dawley rats were fed for 6 weeks with four diets which were based on AIN96G diet and supplemented with Puerariae radix-ethanol extracts to contain isoflavone. The isoflavone contents of four experimental diets were 0 mg, 500 mg, 1,000 mg, 2,000 mg per kg diet, respectively (control, P0.05%, P0.1%, P0.2%). Liver and erythrocyte activities of antioxidative enzyme such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GSHpx) were measured. Also, plasma and liver malondialdehyde (MDA) concentrations, liver glutathione (GSH) and oxidized glutathione (GSSG) concentrations were measured. The total isoflavone content of Puerariae radix-ethanol extract was 3067.6 mg per 100 g extract and the content of puerarin was 2557.4 mg per 100 g extract. The erythrocyte activities of GSH-Px and catalase were higher in group P0.1% and P0.2%. But SOD activity of erythocyte did not show any difference by the Puerariae radix-ethanol extract supplementation in diet. The activity of SOD in liver increased significantly by the supplementation of extract, showing highest level in P0.1% group. The liver GSH concentration increased significantly in group of P0.05%, P0.1%, and P0.2% compared with control group (p < 0.05). The GSSG concentration in liver showed no difference by the supplementation of Puerariae radix extract from the control group, except P0.2% group. The plasma MDA concentration did not show any significant differences by the extract supplementation. But the liver MDA concentration decreased by the extract supplementation, showing the lowest level in P0.1% diet group. These results suggest that the supplementation of Puerariae radix-ethanol extract can inhibit lipid peroxidation in liver and enhance the antioxidative defense competence of rats.

Studies of Chinese Crude Drug "Ge-gen". (II). Collection Season and Differences of Components / 日本東洋医学雑誌

In the previous paper, the authors have investigated a Chinese crude drug Ge-gen, the root of Pueraria species of the family Leguminosae, and concluded that it should be collected in the early summer for medical use. This is a report of the differences in the content of the constituents between Ge-gen harvested in the summer and in the winter. Yields of water extract of winter Ge-gens were generally higher, while no significant difference was recognized in the content of starch and flavonoids, such as puerarin and daidzin. Starch content varies widely with the stock, and that of the same stock did not show significant difference between the summer root and the winter one.