Radix Puerariae Research Articles

Effects of Danggui Sini decoction on neuropathic pain: experimental studies and clinical pharmacological significance of inhibiting glial activation and proinflammatory cytokines in the spinal cord .

Neuropathic pain responds poorly to drug treatments. Partial relief is achieved in only about half of the patients. Danggui Sini decoction (DSD), an aqueous extract of <i>Angelica sinensis, Ramulus Cinnamomi,</i> and <i>Radix Puerariae</i>, has been used extensively in China to treat inflammatory and ischemic diseases. The current study examined the putative effects of DSD on neuropathic pain. We used two commonly-used animal models: chronic constriction injury (CCI) and diabetic neuropathy for the study. And we examined effects of DSD on pain response, activation of microglia and astroglia in spinal dorsal horn, and expression of proinflammatory cytokines in the spinal cord. Consecutive intragastric administration of DSD (25-100mg/kg) for 10 days inhibited the mechanical and thermal nociceptive response induced by CCI and diabetes without interfering with the normal pain response. Meanwhile, in both models, DSD inhibited the over-expression of specific markers for microglia (Iba-1) and astroglia (GFAP) activation in the spinal dorsal horn. DSD also reduced the elevated nuclear NF-κB level and inhibited the up-regulation of proinflammatory cytokines, such as IL-6, IL-1β, and TNF-α, in the spinal cord. DSD can alleviate CCI and diabetes-induced neuropathic pain, and its effectiveness might be due to the inhibition of neuroinflammation in the spinal dorsal horn. The anti-inflammation effect of DSD may be related to the suppression of spinal NF-κB activation and/or cytokines expression. .

Association of traditional Chinese medicine therapy and the risk of dementia in patients with hypertension: a nationwide population-based cohort study

BackgroundPatients with hypertension (HTN) reportedly have a higher risk of developing dementia. However, it remains unclear if use of Traditional Chinese Medicine (TCM), the most common form of complementary and alternative medicine, can help lower the risk of dementia for these patients. So the aim of the study was to investigate the effects of TCM on dementia risk among patients with hypertension.MethodsThis longitudinal cohort study used the Taiwanese National Health Insurance Research Database (NHIRD) to identify 143,382 newly diagnosed hypertension patients aged 20–90 years who received treatment between 1998 and 2007. Among them, 52,365 (36.52%) had received TCM after the onset of hypertension (TCM users), and the remaining 91,017 patients (63.48%) were designated as a control group (non-TCM users). All enrollees were followed until the end of 2012 to record the incidence of dementia. A Cox proportional hazards regression model was used to compute the hazard ratio (HR) of dementia in patients who received TCM.ResultsDuring the 15-year follow-up, 3933 TCM users and 10,316 non-TCM users developed dementia, representing an incidence rate of 8.41 and 11.55%, respectively, per 1000 person-years. TCM users had a significantly reduced risk of dementia compared to non-TCM users (adjusted HR = 0.76; 95% confidence interval [CI] = 0.74–0.81). The predominant effect was observed among those treated with TCM longer than 180 days (adjusted HR = 0.65; 95% CI = 0.62–0.69). Among the commonly used TCM products, Tian-Ma-Gou-Teng-Yin, Dan-Shen (Radix Salviae Miltiorrhizae), Chuan-Niu-Xi (Radix Cyathulae), Ge-Gen (Radix Puerariae), Jia-Wei-Xiao-Yao-San, and Jue-Ming-Zi (Semen Cassiae) were significantly associated with a lower risk of dementia.ConclusionsResults from this population-based study support the effects of TCM on reducing dementia risk, which may provide a reference for dementia prevention strategies.

Analysis of chemical constituents in an herbal formula Jitong Ning Tablet

Jitong Ning Tablet (JTNT), a traditional Chinese herbal formula, consists of Eucommia ulmodies oliv, Angelicae pubescentis radix, Aconiti radix cocta, Corydalis yanhusuo w.t. wang, Glycyrrhizae radix et rhizoma, Paeoniae radix rubra and Radix puerariae. It has been demonstrated to show protective effects on ankylosing spondylitis and anti-inflammatory effects. The chemical compositions of JTNT, playing a key role in quality control, remain unknown. In this study, an ultra-performance liquid chromatography combined with quadrupole time of flight mass spectrometry (UPLC-Q–TOF–MS) method in both positive and negative ion mode was established to investigate the chemical constituents of JTNT formula. In total, 162 compounds including flavonoids, triterpenoids, coumarins, alkaloids, phenylpropionic acids, lignans, terpenoids, and organic acids were detected, 152 of which were unambiguously or tentatively identified by comparing their retention times and accurate mass measurement with reference compounds and data in literatures. Our results would benefit quality control and chemical basis for JTNT.

Puerarin provides a neuroprotection against transient cerebral ischemia by attenuating autophagy at the ischemic penumbra in neurons but not in astrocytes

Puerarin is an isoflavone derived from the Chinese medical herb of Radix puerariae (kudzu root), and has been widely used in the treatment for ischemic stroke in China. However, its underlying pharmacological mechanisms are still not understood. This study was to investigate the efficacy of puerarin on autophagy in the ischemic penumbra after cerebral stroke. A model of cerebral stroke in Sprague-Dawley rats was prepared by middle cerebral artery occlusion (MCAO); rats were then randomly divided into 5 groups: MCAO+Pue group (rats were treated with puerarin), MCAO+Pue+Tat-Beclin-1 group (rats were administrated with both puerarin and autophagy inducer Tat-Beclin-1), MCAO+Tat-Beclin-1 group (rats were treated with Tat-Beclin-1), MCAO+saline group (rats were administrated with the same volume of physiological saline), and sham surgery group. The autophagy levels in infarct penumbra were evaluated by western blotting, real-time PCR and immunofluorescence 14days after the insult. Meanwhile, the neurological deficit score, brain water content and infarct volume were assessed. The results illustrated that the cerebral infarct volume, cerebral edema and neurological deficiency were significantly alleviated by puerarin treatment. Western blotting and the quantitative PCR revealed that the autophagy level in the penumbra was markedly reduced by puerarin administration. However, these effects of puerarin could be counteracted by Tat-Beclin-1. Additionally, double immunofluorescence showed that neuronal autophagy was markedly attenuated by puerarin treatment, whereas astrocytic autophagy was only mildly reduced. Our study suggests that puerarin could confer a neuroprotection against cerebral ischemia, and this biological function is associated with attenuating autophagy in neurons but not in astrocytes.

Puerarin attenuates cisplatin-induced rat nephrotoxicity: The involvement of TLR4/NF-κB signaling pathway.

Puerarin was a major isoflavonoid derived from the Chinese medical herb radix puerariae (Gegen). In present study effect of puerarin on cisplatin nephrotoxicity was evaluated. Rat model of nephrotoxicity was established by a single intraperitoneal injection of cisplatin (7mg/kg). Puerarin was administrated through caudal vein injection once per day at the dose of 10mg/kg, 30mg/kg and 50mg/kg. Biochemical assays showed that after cisplatin treatment the serum urea and creatinine increased significantly compared with control (P<0.05). Cisplatin treatment significantly increased xanthine oxidase (XO) activity and malondialdehyde (MDA) formation, and significantly decreased the levels and /or activities of enzymatic and non-enzymatic antioxidants (GSH, GPx, GST, GR, SOD, CAT), in the kidney tissues. Renal levels of TNF-α and IL-6, two important inflammatory cytokines, were also upregulated by cisplatin. Histopathological examination indicated that cisplatin treatment resulted in severe necrosis and degeneration, hyaline casts in the tubules, intertubular hemorrhage, congestion and swelling in glomerulus and leukocytes infiltration in the kidney tissues. Western blot results demonstrated that cisplatin increased TLR4 and NF-κB protein expression in the kidney tissues. However, all these changes induced by cisplatin were significantly attenuated by puerarin treatment in dose-dependent manner, which indicated the renal protective effect of puerarin. Cell culture experiments illustrated that puerarin alone treatment concentration-dependently inhibited COLO205 and HeLa tumor cell growth and dose-dependently promoted the antitumor activity of cisplatin in COLO205 and HeLa tumor cells. The promotion effects might be attributed to suppression of cisplatin-increased NF-κB p65 expression by puerarin. Taken together, findings in this study suggested that puerarin exhibited renal protection against cisplatin nephrotoxicity via inhibiting TLR4/NF-κB signaling, with no inhibition but promotion effect on the antitumor activity of cisplatin. Puerarin might be a promising adjuvant agent for cisplatin chemotherapy.

Enhancement of antioxidant activity of Radix Puerariae and red yeast rice by mixed fermentation with Monascus purpureus.

In this work, a new functional food combined Radix Puerariae and red yeast rice was explored. The pigment intensity, antioxidant activities and the main isoflavones of it were evaluated and compared with traditional red yeast rice and Radix Puerariae. The fermented mixture showed higher contents of isoflavones and pigment intensities than red yeast rice and Radix Puerariae. The DPPH, OH, FRAP and total antioxidant activity results of fermented mixture also showed higher antioxidant potential than those of Radix Puerariae and red yeast rice, owing to the higher pigment intensity and total phenolic contents. It is concluded that the fermented mixture of Radix Puerariae and rice could be widely used as a source of polyphenols with high antioxidative potential, thus introducing numerous health benefits for the consumer.

The Root Extract of Pueraria lobata and Its Main Compound, Puerarin, Prevent Obesity by Increasing the Energy Metabolism in Skeletal Muscle.

Radix Pueraria lobata (RP) has been reported to prevent obesity and improve glucose metabolism; however, the mechanism responsible for these effects has not been elucidated. The mechanism underlying anti-obesity effect of RP was investigated in high-fat diet (HFD) induced obese mice and skeletal muscle cells (C2C12). Five-week-old C5BL/6 mice were fed a HFD containing or not containing RP (100 or 300 mg/kg) or metformin (250 mg/kg) for 16 weeks. RP reduced body weight gain, lipid accumulation in liver, and adipocyte and blood lipid levels. In addition, RP dose-dependently improved hyperglycemia, insulinemia, and glucose tolerance, and prevented the skeletal muscle atrophy induced by HFD. Furthermore, RP increased the peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) expression and phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) in skeletal muscle tissues. RP and its main component, puerarin, increased mitochondrial biogenesis and myotube hypertrophy in C2C12 cells. The present study demonstrates that RP can prevent diet-induced obesity, glucose tolerance, and skeletal muscle atrophy in mouse models of obesity. The mechanism responsible for the effect of RP appears to be related to the upregulation of energy metabolism in skeletal muscle, which at the molecular level may be associated with PGC-1α and AMPK activation.

Research progress of puerarin's antitumor mechanism

Puerarin is a kind of isoflavone compounds extracted from Chinese herbal medicine radix puerariae, and have antitumor effect which can significantly inhibit lung, stomach, colon cancer, liver cancer, lymphoma cell growth and induce its apoptosis.Related studies had shown that it play a role in anti-tumor mainly by blocking the cell cycle, inducing apoptosis, interventing mitochondrial regulation, injuring mitochondrial cells, and influencing tumor apoptosis signaling pathways et al. Key words: Puerarin; Tumor; Mechanism

Fermented Chinese formula Shuan-Tong-Ling attenuates ischemic stroke by inhibiting inflammation and apoptosis.

The fermented Chinese formula Shuan-Tong-Ling is composed of radix puerariae (Gegen), salvia miltiorrhiza (Danshen), radix curcuma (Jianghuang), hawthorn (Shanzha), salvia chinensis (Shijianchuan), sinapis alba (Baijiezi), astragalus (Huangqi), panax japonicas (Zhujieshen), atractylodes macrocephala koidz (Baizhu), radix paeoniae alba (Baishao), bupleurum (Chaihu), chrysanthemum (Juhua), rhizoma cyperi (Xiangfu) and gastrodin (Tianma), whose aqueous extract was fermented with lactobacillus, bacillus aceticus and saccharomycetes. Shuan-Tong-Ling is a formula used to treat brain diseases including ischemic stroke, migraine, and vascular dementia. Shuan-Tong-Ling attenuated H2O2-induced oxidative stress in rat microvascular endothelial cells. However, the potential mechanism involved in these effects is poorly understood. Rats were intragastrically treated with 5.7 or 17.2 mL/kg Shuan-Tong-Ling for 7 days before middle cerebral artery occlusion was induced. The results indicated Shuan-Tong-Ling had a cerebral protective effect by reducing infarct volume and increasing neurological scores. Shuan-Tong-Ling also decreased tumor necrosis factor-α and interleukin-1β levels in the hippocampus on the ischemic side. In addition, Shuan-Tong-Ling upregulated the expression of SIRT1 and Bcl-2 and downregulated the expression of acetylated-protein 53 and Bax. Injection of 5 mg/kg silent information regulator 1 (SIRT1) inhibitor EX527 into the subarachnoid space once every 2 days, four times, reversed the above changes. These results demonstrate that Shuan-Tong-Ling might benefit cerebral ischemia/reperfusion injury by reducing inflammation and apoptosis through activation of the SIRT1 signaling pathway.

A novel herbal treatment reduces depressive-like behaviors and increases brain-derived neurotrophic factor levels in the brain of type 2 diabetic rats.

BackgroundRadix Puerariae and hawthorn fruit have been demonstrated to treat diabetes. They offer potential benefits for preventing depression in diabetes.ObjectiveThe aim of this study was to investigate whether the combination of Radix Puerariae and hawthorn fruit (CRPHF) could prevent depression in a diabetic rat model generated by feeding the rats with a high-fat diet and a low-dose streptozotocin (STZ).MethodsThe CRPHF was provided by the Shanghai Chinese Traditional Medical University. Twenty-four rats were randomly divided into four groups: normal control, normal-given-CRPHF (NC), diabetic control, and diabetic-given-CRPHF (DC) groups. The type 2 diabetic model was created by feeding the rats with a high-fat diet for 4 weeks followed by injection of 25 mg/kg STZ. CRPHF was given at 2 g/kg/d to the rats of NC and DC groups by intragastric gavage daily for 4 weeks after the type 2 diabetic model was successfully created. Body weight, random blood glucose (RBG), oral glucose tolerance test, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured during the study. Depressive-like behavior was evaluated at the end of the treatment by using the open field test (OFT), the elevated plus-maze test (EPMT), locomotor activity test (LAT), and forced swimming test (FST). Levels of extracellular signal-regulated protein kinase (ERK) and brain-derived neurotrophic factor (BDNF) in the prefrontal cortex were evaluated by using Western blot.Results1) CRPHF reduced RBG and improved glucose tolerance in diabetic rats; 2) CRPHF reduced TC and TG but did not significantly change HDL-C or LDL-C in diabetic rats; 3) CRPHF reversed the loss in body weights observed in diabetic rats; 4) CRPHF reduced depressive-like behavior as measured by OFT, EPMT, LAT, and FST; 5) BDNF was upregulated, and ERK was activated in the prefrontal cortex of diabetic rats treated with CRPHF.ConclusionCRPHF has the potential of preventing depression in patients with diabetes.

Safety and effectiveness of different herbal medicine dosage of Gegen Qinlian Decoction in Chinese patients with type 2 diabetes: a double-blind, two-part, randomised controlled trial

BackgroundGegen Qinlian Decoction, a Chinese herbal medicine, has been shown to be effective and safe for patients with type 2 diabetes, yet the roles of its main components, Radix Puerariae and Coptis chinensis, in diabetes is uncertain. We aimed to observe the relationship between dosage-changing of these herbal medicines of Gegen Qinlian Decoction and effectiveness in patients with type 2 diabetes. MethodsWe did a randomised, double-blind, two-part, parallel-dose controlled clinical trial. Patients with type 2 diabetes inadequately controlled by diet and exercise from Guang'anmen Hospital were enrolled in the phase 1 trial. Patients who had been treated for diabetes within 1 month were excluded from the trial. Patients were randomly allocated (1:1:1) via a computer-generated randomisation sequence to receive three different doses (24 g, 72 g, and 120 g) of Radix Puerariae of Gegen Qinlian Decoction for 12 consecutive weeks. Patients and clinicians were masked to group assignment. The primary outcomes were changes from baseline in HbA1c, fasting plasma glucose, and 2 h glucose after oral glucose tolerance test at week 12 in each group, and the outcomes were analysed in the per-protocol population who complied with the protocol and had no violations. Secondary endpoints included concentrations of fasting insulin and lipids, bodyweight, BMI, and waist circumference. If the relationship between dosage-changing in Radix Puerariae of Gegen Qinlian Decoction and HbA1c-decreasing was null, the new patients with diabetes were enrolled in the phase 2 trial, which had patients randomly allocated (1:1:1) to receive three different doses Coptis chinensis of Gegen Qinlian Decoction for 12 consecutive weeks. The efficacy evaluation was identical to the phase 1 trial. This study is registered with ClinicalTrials.gov, number NCT01219803. FindingsBetween Aug 8, 2011, and April 23, 2012, we enrolled 120 patients into the phase 1 trial. 97 patients were included in the per-protocol analysis (32 for high dose, 30 for middle dose, and 35 for low dose Radix Puerariae of Gegen Qinlian Decoction). At week 12, No significant difference among the groups was reported compared with week 0: the mean change in HbA1c was −0·58% in the high-dose group (SD 0·87), −0·28% in the middle-dose group (1·17), and −0·55% in the low-dose group (0·85; the difference between groups p=0·52, 95% CI −0·53 to 0·55). In the phase 2 trial, we enrolled another 122 patients between Nov 14, 2012, and Aug 29, 2013, and 103 of them were included in the per-protocol analysis. Compared with the HbA1c level from week 0, the mean change was −0·75% in the high-dose group (SD 0·82), −0·34% in the middle-dose group (0·71), and −0·26% in the low-dose group (0·79) at week 12. There was a significant HbA1c reduction in the groups after 12 weeks (p=0·0492, 95% CI −0·78 to −0·02). No cases of severe hypoglycaemia were reported in either trial. InterpretationChanging the dose of Coptis chinensis of Gegen Qinlian Decoction has a dose–effect relationship with glycaemic control. Coptis chinensis is the major drug of Gegen Qinlian Decoction for treating patients with type 2 diabetes. FundingNational Basic Research Program of China (973 Program).

Puerarin, an isoflavone compound extracted from Gegen (Radix Puerariae Lobatae), modulates sclera remodeling caused by extremely low frequency electromagnetic fields

OBJECTIVETo evaluate the protective effect of puerarin [an isoflavone compound extracted from Gegen (Radix Puerariae Lobatae)] in scleral remodeling induced by extremely low frequency electromagnetic fields (ELF-EMFs). METHODSHuman fetal scleral fibroblasts (HFSFs) were divided into 5 groups: (1) untreated controls; (2) cells treated with ELF-EMFs; (3) cells treated with ELF-EMFs and puerarin 0.1 μM; (4) cells treated with ELF-EMFs and puerarin 1 μM; (5) cells treated with ELF-EMFs and puerarin 10 μM. Cell proliferation activity was measured by the cell-counting kit-8 assay. Matrix metalloproteinase-2 (MMP-2) activity was measured by gelatin enzymography. MMP-2 and collagenI(COL1A1) mRNA, protein expression were measured by Real-Time polymerase chain reaction, Western blot analysis, respectively. RESULTSPuerarin reduced the inhibition in cell proliferation, MMP-2 activity, mRNA, protein expression of HFSFs exposed to ELF-EMFs and enhanced the COL1A1 mRNA and protein expression. CONCLUSIONPuerarin was found to participate in the matrix remodeling process. It might be a potential agent for the treatment of extracellular matrix degradation of sclera associated with ocular conditions.

Puerarin ameliorates heat stress–induced oxidative damage and apoptosis in bovine Sertoli cells by suppressing ROS production and upregulating Hsp72 expression

Puerarin, a bioactive isoflavone glucoside extracted from radix Puerariae, has been proven to possess many biological activities. However, the role of puerarin in protecting bovine Sertoli cells (bSCs) under heat stress conditions remains to be clarified. The present study aimed to explore the possible protective mechanism of puerarin for primary cultured bSCs subjected to heat stress. Bovine Sertoli cells were treated with 15 μM of puerarin before they were exposed to 42 °C for 1 hour. The dose of puerarin (15 μM) was determined on the basis of cell viability. The results showed that puerarin treatment suppressed the production of reactive oxygen species and decreased the oxidative damage of the bSCs subjected to heat stress, as indicated by changes in superoxide dismutase, catalase, and glutathione peroxidase activities and malondialdehyde content. Moreover, puerarin treatment also suppressed the initiation of mitochondria-dependent apoptotic pathway, as revealed by changes in Bax to Bcl-2 ratio, mitochondrial membrane potential, cytochrome C release, caspase-3 activation, and apoptotic rate compared with the heat stress group. In addition, puerarin treatment increased Hsp72 expression in the bSCs with no apparent cellular cytotoxicity compared with the control group. Furthermore, increased Hsp72 was detected in the heat stress plus puerarin group compared with the heat stress group. In conclusion, puerarin attenuates heat stress–induced oxidative damage and apoptosis of bSCs by suppressing reactive oxygen species production and upregulating Hsp72 expression.

Selenium speciation in radix puerariae using ultrasonic assisted extraction combined with reversed phase high performance liquid chromatography-inductively coupled plasma-mass spectrometry after magnetic solid-phase extraction with 5-sulfosalicylic acid functionalized magnetic nanoparticles

A new method for determination of selenium species in radix puerariae was described. The method consists of sample enrichment with 5-sulfosalicylic acid (SSA)-functionalized silica-coated magnetic nanoparticles (SMNPs), high performance liquid chromatography (HPLC) separation, and online detection using inductively coupled plasma mass spectrometry (ICP-MS). The selenium species were extracted using ultrasonic extraction system with a mixture of protease K and lipase. The SSA-SMNPs were used to enrich trace amounts of selenite [Se(IV)], selenate [Se(VI)], selenomethionine (SeMet), and selenocystine (SeCys2) from lower selenium containing samples. Under the optimal conditions, the limits of detection (3σ) for SeCys2, Se(IV), SeMet and Se(VI) were observed as 0.0023, 0.0015, 0.0043, and 0.0016ngmL−1, respectively. The RSD values (n=6) of method for intraday were observed between 0.5% and 0.9%. The RSD values of method for interday were less than 1.3%. The linear concentration ranges for SeCys2, Se(IV), SeMet and Se(VI) were 0.008–1000, 0.005–200, 0.015–500 and 0.006–200ngmL−1, respectively. The detection limits of this method were improved by 10 times due to the enrichment with the SSA-SMNP extraction. The contents of SeCys2, Se(IV), SeMet, and Se(VI) in radix puerariae were determined as 0.0140, 0.171, 0.0178, and 0.0344μgg−1, respectively. The recoveries were in the range of 95.6%–99.4% and the RSDs (n=6) of recoveries were less than 1.5%.

Puerarin Exerts an Antiinflammatory Effect by Inhibiting NF-kB and MAPK Activation in Staphylococcus aureus-Induced Mastitis.

Mastitis is defined as the inflammation of the mammary gland. There is generally no effective treatment for mastitis in animals. Puerarin, extracted from Radix puerariae, has been proven to possess many biological activities. The present study aims to reveal the potential mechanism that is responsible for the antiinflammatory action of puerarin in Staphylococcus aureus (S. aureus)-induced mastitis in mice. Histopathological changes showed that puerarin ameliorated the inflammatory injury induced by S. aureus. Quantitative real-time polymerase chain reaction and ELISA analysis indicated that puerarin not only suppressed the production of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 but also promoted the secretion of IL-10. Toll-like receptor 2 (TLR2) is important in the immune defense against S. aureus infection. Research in molecular biology has shown that the expression of TLR2 was inhibited with administration of puerarin. Further studies were performed on NF-kB and mitogen-activated protein kinase signaling pathways using western blot. The results demonstrated that puerarin suppressed phosphorylated IkBα, p65, p38, extracellular signal-regulated kinase 1and 2 (ERK), and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. All of the results suggested that puerarin may be a potential therapy for treating mastitis. Copyright © 2016 John Wiley & Sons, Ltd.

Analysis of Eight Isoflavones in Radix Puerariae by MEEKC: Comparison on Three Different Oil Phases

In this study, three different oil phases include 1-butyl-1-methylpyrrolidinium bis(trifluoromethanesulfonyl)imide (BMPy[NTf2]), 1-butyl-3-methylimidazolium hexafluorophosphate (BMImPF6) and n-octane were compared for the MEEKC analysis (SDS as surfactant and n-butanol as co-surfactant) of eight isoflavones from pueraria. The investigated isoflavones can be well separated by all of those three microemulsion systems after careful optimization, and the MEEKC with n-octane as the oil phase was the best choice (good symmetry and high resolutions of peaks with short analysis time) for the analysis. The optimum conditions of MEEKC method were as follows: 70 mM SDS, 0.7 M n-butanol and 0.5% (w/v) n-octane in 10 mM sodium tetraborate (STB) at pH 8.5, applied voltage was 23 kV and cassette temperature was set at 30°C. And then the developed method was fully validated (limit of detection, limit of quantification, intraday precision, interday precision and recovery) and successfully applied to determine the eight analytes in three Radix Puerariae samples. In addition, although the MEEKC with classic oil phase (n-octane) showed better results for isoflavones analysis in this study, the MEEKC with ionic liquids (BMPy[NTf2] and BMImPF6) also showed great separation potential for analytes, which may be further applied in the analysis of other natural products.

Screening and isolation of potential lactate dehydrogenase inhibitors from five Chinese medicinal herbs: Soybean, Radix pueraria, Flos pueraria, Rhizoma belamcandae, and Radix astragali.

Stroke is among the leading causes of death and severe disability worldwide. Flavonoids have been extensively used in the treatment of ischemic stroke by reducing lactate dehydrogenase levels and thereby enhancing blood perfusion to the ischemic region. Here, we used ultrafiltration high-performance liquid chromatography coupled with diode array detection and mass spectrometry for the rapid screening and identification of flavonoids from five Chinese medicinal herbs: soybean, Radix pueraria, Flos pueraria, Rhizoma belamcandae, and Radix astragali. Using PC12 cells as a suitable in vitro model of toxicity, cell viability was quantitated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The results showed that the extracts of soybean and the six major components, namely, acetyldaidzin, malonylgenistin, daidiain, glycitin, genistin, and acetylcitin; the extract of R. pueraria and its main component daidzein; the extract of F. pueraria and its three major components, tectorigenin, tectoridin, and tectorigenin-7-O-xylosylglucosid; and the extract of R. belamcandae and its main component, tectoridin, were strong lactate dehydrogenase inhibitors. Also, the components of R. astragali showed no bioactivity. These findings indicate that the ultrafltration high-performance liquid chromatography coupled with diode array detection and mass spectrometry method could be utilized in rapid screening and separation of bioactive compounds from a complex matrix.

NMR Study on the Inclusion Complexes of β-Cyclodextrin with Isoflavones.

The structure of the inclusion complexes of β-cyclodextrin (β-CD) with daidzein and daidzin in D2O were investigated using NMR spectroscopy. For the β-CD and daidzein system, two types of 1:1 complexes were formed with the daidzein deeply inserted into the CD cavity with different orientations. For the β-CD/daidzin system, a 1:1 complex was formed with the flavonoid part of daidzin entering the CD cavity from the wide rim. The inclusion complexes determined by NMR were constructed using molecular docking. Furthermore, the mixture of puerarin, daidzein and daidzin, which are the major isoflavonoid components present in Radix puerariae, was analyzed by diffusion-ordered spectroscopy (DOSY) alone and upon addition of β-CD in order to mimic chromatographic conditions and compare their binding affinities.

Radix Puerariae lobatae (Gegen) suppresses the anticoagulation effect of warfarin: a pharmacokinetic and pharmacodynamics study.

BackgroundRadix Salvia miltiorrhiza (Danshen) and Radix Puerariae lobatae (Gegen) are used in Traditional Chinese Medicine to treat cardiovascular diseases. However, adverse herb-drug interactions were observed between warfarin and herbal remedies containing Danshen and Gegen. This study aims to investigate the pharmacokinetic and pharmacodynamic interactions between warfarin and the different components found in Danshen and Gegen.MethodsSixty Sprague–Dawley rats were used to investigate the effects of warfarin (0.2 mg/kg), Danshen (240 or 480 mg/kg) and Gegen (240 or 480 mg/kg) both in isolation and combination. The rats in the warfarin and Danshen/Gegen combination groups were given an oral dose of Danshen or Gegen 2 h after being given an oral dose of warfarin. After five consecutive days of treatment, the pharmacokinetic interactions between Danshen/Gegen and warfarin were investigated by simultaneously monitoring and comparing the cytochrome P450 (CYP) activities, mRNA and protein expression levels in the livers of the rats from the different treatment groups. The pharmacodynamic interactions were evaluated by monitoring and comparing the vitamin K epoxide reductase (VKOR) activities, mRNA and protein expression levels in the livers of rats from the different groups, as well as the thrombomodulin (TM) activities, mRNA and protein in the lungs of these animals. The rat plasma soluble thrombomodulin concentrations of the different treatment groups were also evaluated. Microsomes incubation, Real Time-Polymerase Chain Reaction and Western blot was applied respectively to study the activity, mRNA expression and protein expression of CYP, VKOR and TM.ResultsThe activities and expression levels of the CYP and VKOR enzymes in the warfarin-Gegen combination groups increased by nearly 30 % (P = 0.02) compared with the warfarin-alone group, whereas those of TM decreased by almost 25 % (P = 0.02). The administration of Danshen did not lead to any changes in the activities or the expression levels of the CYP, VKOR or TM enzymes compared with those of the control group. Gegen induced several warfarin-metabolizing CYP enzymes and neutralized the effects of warfarin towards VKOR and TM.ConclusionGegen, rather than Danshen at the same tested dosage, offsets the anticoagulant effects of warfarin by accelerating the phase I liver metabolism of warfarin, as well as increasing the activity, mRNA and protein expression of VKOR while decreasing those of TM.Electronic supplementary materialThe online version of this article (doi:10.1186/s13020-016-0078-9) contains supplementary material, which is available to authorized users.

Pharmacokinetic profiles of the five isoflavonoids from Pueraria lobata roots in the CSF and plasma of rats

Ethnopharmacological relevanceTraditional Chinese medicine Radix Puerariae, the roots of Pueraria lobata (Wild.) Ohwi., has been widely used for the treatment of cardiovascular and cerebrovascular diseases in China for centuries. Isoflavonoids are believed the active components of this herb. Aim of this studyThe present study aims to investigate the brain penetration and pharmacokinetics of five active isoflavonoids in the ventricular CSF and plasma of rats after intravenous administration of a Pueraria isoflavonoids (PIF) extract, to better understand the active components of this herb for neuro-activities. Material and methodsUnder anesthesia condition, SD rats (n=6) were successively suffered two surgeries for implanting cannulas at lateral ventricle and right jugular vein for brain microdialysis and blood collection, respectively. After recovery, the rats received intravenous dose of PIF at 80mg/kg and the concentrations of puerarin (PU), 3′-methoxypuerarin (MPU), 3′-hydroxypuerarin (HPU), daidzein (DA) and daidzein-8-C-apiosyl-(1-6)-glycoside (DAC) in the ventricular dialysate and plasma samples were determined using a ultra-fast liquid chromatography tandem mass spectrometry method. ResultsComplete concentration versus time profiles of the five components in plasma and four components except for HPU in ventricular CSF were obtained. After dosing, the average C0 values of PU, MPU, DA, DAC and HPU in plasma were estimated 6.53, 13.72, 1.54, 15.84 and 86.07µg/mL, and PU, MPU, DA and DAC were rapidly penetrated to the brain and reached to their Cmax of 521.52, 415.00, 74.34 and 380.03ng/mL in CSF at about 0.5–0.8h, respectively. The elimination t1/2 of PU, DA and DAC in CSF and plasma were no significant difference, while the t1/2 of MPU in ventricular CSF was longer than that in plasma which may attributable to the different physiological environment of central and peripheral compartments. The brain penetration index (AUCCSF/AUCplasma) was found to be about 9.29, 7.25, 11.96, and 4.21% for PU, MPU, DA, and DAC respectively. ConclusionPU, MPU, DA, DAC can quickly penetrate to the brain through the blood brain barrier (BBB) and might be responsible for the neuro-pharmacological activities of P. lobata.