Evaluation of Relationship of Pueraria lobata Quality from Different Producing Areas Based on "Climate Factor-component Content-antioxidant Capacity" / 中国实验方剂学杂志

ObjectiveThe correlation of Pueraria lobata producing areas， climate factors， total flavonoids of P. lobata， polysaccharide content of P. lobata， and antioxidant activity of P.lobata for medicinal application was analyzed， and the relationship between climate factors and the formation of P. lobata quality was evaluated. MethodThe scavenging rates of 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl（DPPH） and 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)（ABTS） radicals by total flavonoids and polysaccharides of P. lobata were detected， and the correlation between the contents of each component and the information of producing areas and climate factors was analyzed. ResultThe ABTS+ scavenging rate by total flavonoids of P.lobata was negatively correlated with altitude （P<0.05） and positively correlated with annual sunshine hours （P<0.05）. The altitude was positively correlated with the total flavonoid content， while the annual sunshine hours were negatively correlated with the total flavonoid content. There was a negative correlation between total flavonoid content and ABTS+ scavenging rate by total flavonoids. In other words， lower altitude and longer annual sunshine hours indicated lower total flavonoid content and higher ABTS+ scavenging rate by total flavonoids. The ABTS+ scavenging rate by polysaccharides of P. lobata was negatively correlated with the frost-free period （P<0.05） and the mean temperature in July （P<0.01）. There was a positive correlation between the polysaccharide content of P. lobata and the frost-free period. The mean temperature in July was positively correlated with the polysaccharide content of P. lobata （P<0.05）. The polysaccharide content of P. lobata was negatively correlated with the ABTS+ scavenging rate by polysaccharides of P. lobata. In other words， a shorter frost-free period in the producing area and lower mean temperature in July indicated lower polysaccharide content of P. lobata and higher ABTS+ scavenging rate by polysaccharides of P. lobata. The mean temperature in July was significantly correlated with the contents of total flavonoids and polysaccharides in P. lobata samples （P<0.05）. The lower mean temperature in July was often accompanied by lower total flavonoid content of P. lobata， lower polysaccharide content of Pueraria lobata， and stronger antioxidant activity of P. lobata samples. ConclusionThe ability of P. lobata to remove ABTS+ is stronger than that of DPPH+. There is a significant correlation between climate factors， content， and antioxidant capacity in each producing area. Further research on the internal law of the formation of medicinal active components of P. lobata induced by core climate factors will provide a scientific basis for revealing the formation mechanism of genuine P. lobata and the subsequent control of P. lobata quality according to the environment of producing areas.

Crystallization transformation of amorphous extracts of traditional Chinese medicine and its effect on dissolution behavior — Taking total flavonoids from Pueraria lobata as an example / 中国药科大学学报

@#In order to guarantee the quality of traditional Chinese medicines (TCMs), the crystallization transformation of complex extracts of TCMs and the influence of solid form on their physicochemical properties were studied.The extract of total flavonoids from Pueraria lobata was taken as a model.Crystallization transformation happened when lofting under different conditions, and the intrinsic dissolution tests were carried out.It was found that humidity was the key factor to induce crystallization of total flavonoids from Pueraria lobata.The greater the wettability was, the more the crystallization was.The dissolution rate of total flavonoids from Pueraria lobata with the most crystallization amount significantly decreased by 96.51% compared to the sample without crystallization.After further simulating the preparation process of total flavonoids from Pueraria lobata, it was found that the wet granulation process with introduced water would also lead to crystallization and reduced dissolution rate.As for all crystallization samples, there was an inversely proportional relationship between the dissolution rates and the amount of crystallization.The risk of crystallization existed both in the storage and preparation process of TCM extracts.Crystallization would significantly affect the dissolution rate, and thus the quality of TCM products.In this study, the crystallization transformation of amorphous complex TCM extracts was discovered, and the effect of the crystallization transformation on its dissolution behavior was systematically studied, which provides a new research idea for assuring the quality of TCM products and promoting the improvement of TCM preparation level.

Nanocrystals self-stabilized Pickering emulsion loaded with active components of Tongmai prescription: preparation, characterization and evaluation by Caco-2 cell model / 药学学报

It is of great significance to apply the nanocrystals self-stabilized Pickering emulsion (NSSPE) to traditional Chinese medicine (TCM) compounds, and to study the effect of NSSPE on the oral absorption of various components with different solubility and permeability. In the study, NSSPE of Tongmai prescription was prepared by the high pressure homogenization method with nanocrystals of main active components (puerarin, ferulic acid, salvianolic acid B and tanshinone IIA) of Tongmai prescription as solid particle stabilizers and a mixture of Ligusticum chuanxiong essential oil and Labrafil M 1944 CS as oil phase. The NSSPE had better physical stability than nanocrystals suspension and blank emulsion. The adsorption of nanocrystals on the surface of oil droplets was confirmed by scanning electron microscopy and fluorescence microscopy. The surface adsorption rates of puerarin, ferulic acid, salvianolic acid B and tanshinone ⅡA in NSSPE were 15.40% ± 3.19%, 15.39% ± 5.07%, 10.97% ± 3.70% and 31.51% ± 1.60%, respectively. When solid active components were prepared into nanocrystals suspension, the cellular uptake and transport across Caco-2 cells were increased significantly for puerarin and tanshinone IIA. The uptake rates of ferulic acid, ligustilide and tanshinone IIA in NSSPE were further increased compared with the physical mixture of nanocrystals suspension and oil, and the transports of ligustilide and tanshinone IIA were also significantly improved. The main absorption mechanisms of NSSPE were passive diffusion and caveolin-mediated endocytosis, which were determined mainly by the microstructure of NSSPE. In conclusion, NSSPE could be applied to complicated TCM. The "micro" and "nano" synergistic microstructure with drug nanocrystals adsorbed on the surface of micron-sized oil droplets could not only improve the physical stability of NSSPE, but also promote the absorption of various components in NSSPE, which made NSSPE a promising oral drug delivery system for TCM.

Effects of pueraria lobata combined with high-intensity intermittent training on skeletal muscle quality and gene expression / Efeitos da pueraria lobata combinados com treino intermitente de alta intensidade na qualidade músculoesquelética e expressão genética / Efecto de pueraria lobata y entrenamiento intermitente de alta intensidad sobre la calidad del músculo esquelético y la expresión génica

ABSTRACT Extraction of effective components from Pueraria lobata has important value for skeletal muscle quality and gene expression. The improvement effect of traditional high-intensity intermittent training on skeletal muscle has not been obvious, and it is difficult to guarantee the properties of some volatiles. Based on this, this paper analyzes the effect of high-intensity intermittent training on skeletal muscle quality and gene expression in Pueraria lobata. Based on a brief summary of extraction of Pueraria lobata, status of research on the pharmaceutical components of Pueraria lobata was summarized. Different specimens of Pueraria lobata were selected as research objects, and the process of high-intensity intermittent training was designed. High-intensity intermittent training, solvent extraction and water solvent extraction were combined together to design the fixed-bed continuous extraction scheme. According to the influence of Pueraria lobata on skeletal muscle quality, the influence of intermittent training on skeletal muscle quality was analyzed. The extraction results showed that Pueraria lobata combined with high-intensity intermittent training can effectively improve the content of skeletal muscle and ensure the effective expression of skeletal muscle gene.

RESUMO A extração de componentes eficazes da Pueraria lobata tem importante valor para a qualidade músculoesquelética e para a expressão genética. O efeito da melhoria do tradicional treinamento intervalado de alta intensidade na estrutura músculoesquelética não tem sido óbvio, e é difícil garantir as propriedades de alguns voláteis. Com base nisso, este estudo analisa o efeito do treinamento intervalado de alta intensidade na qualidade músculoesquelética e na expressão genética na Pueraria lobata. Com base num breve resumo da extração da Pueraria lobata, resumiu-se o andamento das pesquisas sobre os componentes farmacêuticos da Pueraria lobata. Diferentes amostras de Pueraria lobata foram selecionadas como objeto de pesquisa, e formulou-se o processo do treinamento intervalado de alta intensidade. O treinamento intervalado de alta intensidade, a extração de solventes e a extração de solventes à base de água foram combinadas para conceber o sistema de extração contínua de leito fixo. De acordo com a influência da Pueraria lobata na qualidade músculoesquelética, analisou-se a influência do treino intervalado na qualidade músculoesquelética. Os resultados da extração mostraram que a Pueraria lobata, combinada com treino intervalado de alta intensidade, pode melhorar, de maneira eficaz, o teor músculoesquelético e garantir a expressão eficaz da expressão genética do músculoesquelético.

RESUMEN La extracción de componentes eficaces de la Pueraria lobata tiene un importante valor para la calidad músculoesquelética y para la expresión genética. El efecto de la mejora del tradicional entrenamiento intercalado de alta intensidad en la estructura músculoesquelética no ha sido obvio, y es difícil garantizar las propriedades de algunos volátiles. Basándose en eso, este estudio analiza el efecto del entrenamiento intercalado de alta intensidad en la calidad músculoesquelética y en la expresión genética en la Pueraria lobata. Basándose en un breve resumen de la extracción de la Pueraria lobata, se resumió el andamiento de las investigaciones sobre los componentes farmacéuticos de la Pueraria lobata. Diferentes muestras de Pueraria lobata fueron seleccionadas como objeto de investigación, y se formuló el proceso del entrenamiento intercalado de alta intensidad. El entrenamiento intercalado de alta intensidad, la extracción de solventes y la extracción de solventes a base de agua fueron combinadas para concebir el sistema de extracción continua de lecho fijo. De acuerdo con la influencia de la Pueraria lobata en la calidad músculoesquelética, se analizó la influencia del entrenamiento intercalado en la calidad músculoesquelética. Los resultados de la extracción mostraron que la Pueraria lobata, combinada con entrenamiento intercalado de alta intensidad, puede mejorar, de manera eficaz, el tenor músculoesquelético y garantizar la expresión eficaz de la expresión genética del músculoesquelético.

Difference Analysis of Chemical Constituents in Different Varieties of Puerariae Flos by UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective:To compare the chemical constituents of Puerariae Flos from three different varieties of <italic>Pueraria montana</italic> var. <italic>lobata</italic>， <italic>P. montana</italic> var. <italic>thomsonii</italic> and <italic>P</italic>. <italic>montana</italic> var<italic>. montana</italic>. Method:Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS） was used with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-20 min， 10%-30%B； 20-30 min， 30%-55%B； 30-35 min， 55%-95%B； 35-37 min， 95%B； 37-40 min， 95%-10%B）， the flow rate was 0.25 mL·min^-1. Electrospray ionization （ESI） was used to scan and collect MS data in positive and negative ion modes with scanning range of <italic>m</italic>/<italic>z</italic> 50-1 500. The chemical components from different sources of Puerariae Flos were identified in combination with the chemical composition database and literature information. After the obtained data were normalized by MarkerView^TM 1.2.1， they were imported into SICMA-P 14.1 software for principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） to select the main differentiated components among the three different varieties. Result:A total of 35 compounds were identified from three different varieties of Puerariae Flos， including 22 isoflavones， 6 flavonoids and 7 saponins. The flowers of <italic>P</italic>. <italic>lobata</italic>， <italic>P. montana</italic> var. <italic>thomsonii</italic> and <italic>P</italic>. <italic>montana</italic> var<italic>. montana</italic> contained 32， 35， 33 compounds， respectively. And 18 differential compounds were screened under the positive and negative ion modes， including kakkalide， tectoridin， 6″-<italic>O</italic>-xylosyl-tectoridin， 4'-methyltectorigenin-7-glucoside， glycitin， 6″-<italic>O</italic>-xylosyl-glycitin， irisolidone， kaikasaponin Ⅲ， 6″-<italic>O</italic>-malonylglycitin， kakkalidone， tectorigenin， rutin， soyasaponin BB， vitexin， biochanin A， genistin， kakkatin， azukisaponin Ⅱ. Conclusion:This research is the first to systematically study the chemical constituents of the flower of <italic>P</italic>. <italic>montana</italic> var<italic>. montana</italic>， although the flower of <italic>P</italic>. <italic>montana</italic> var<italic>. montana</italic> is used as adulterants， it has high contents of tectoridin and 6″-<italic>O</italic>-xylosyl-tectoridin， which has great potential for development. The efficacy components such as kakkalide and tectoridin in Puerariae Flos from the three sources of varieties are obviously different， and it is necessary to carefully consider the application of these three varieties as Puerariae Flos.

Effective Component Screening in Antioxidant Active Fraction of Pueraria lobata by Fingerprint Combined with Chemometrics / 中国药房

OBJECTIVE：To screen the effective compo nent in antioxi dant active fraction of Pueraria lobata . METHODS ：The antioxidant active fraction sample （S1-S20） of 20 batches of P. lobata were prepared. HPLC method was adopted. The determination was performed on SepaxBio-C 18 column with mobile phase consisted of methanol-water （gradient elution ）at the flow rate of 0.6 mL/min. The column temperature was set at 25 ℃，and detection wavelength was set at 250 nm. HPLC fingerprints of 20 batches of P. lobata were established by the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition），and common peaks were identified. Cluster analysis ，principal component analysis （PCA）and orthogonal partial least squares discriminant analysis （OPLS-DA）were used to screen the effective components in antioxidant active fraction of P. lobata . RESULTS：There were 18 common peaks in HPLC fingerprints of 20 batches of antioxidant active fraction in P. lobata ，and the similarity was more than 0.99. Eight common peaks were identified ，which were 3′-hydroxypuerarin（peak 2），puerarin（peak 3）， 3′-methoxypuerarin（peak 4），daidzein（peak 5），genistein（peak 7），formononetin（peak 11），daidzein（peak 13）and genistein （peak 16）. The results of cluster analysis and PCA analysis showed that samples S 1，S3，S4，S6，S8，S18 and S 19 were clustered into one category ，and samples S 2，S5，S7，S9-S17 and S 20 were clustered into one category ；peak 2，peak 3，peak 10，peak 11 and peak 13 had great influence on principal component 1；peak 8 and peak 9 had great influence on principal component 2. OPLS-DA analysis showed that peak 4，peak 3，peak 2，peak 16，peak 13 and peak 11 had great influence on the quality of antioxidant active fraction of P. lobata . CONCLUSIONS ： HPLC fingerprint for active fraction of P. lobata is established in the study and 8 components are identified ；among them ， com puerarin，3′-hydroxypuerarin，daidzein and formononetin maybe the material basis of antioxidant fraction of P. lobata .

Pueraria lobate advances in molecular pharmacognosy / 中国中药杂志

Molecular pharmacognosy is a science of classification and identification, cultivation and protection, and production of active ingredients of graduated drugs at the molecular level. The proposal of molecular pharmacognosy allows the research of crude drugs to advance from the microscopic level to the genetic level. Pueraria lobata root, as a medicinal and edible plant, has high application value and economic value. There are many varieties that are easy to cause confusion, and it is not easy to distinguish and identify according to traditional identification methods. Moreover, the research of P. lobate root at the genetic level is still relatively shallow. the study received extensive attention of scholars. This article reviews recent research on molecular identification of P. lobate, transcriptome sequencing, cloning and synthesis of functional genes of P. lobate root in recent years in order to provide references for further promoting the development and utilization of P. lobate root and its active ingredients.

Research advances in chemical constituents and pharmacological activities of Pueraria genus / 中国中药杂志

Kudzu plants in the subfamily sphenoideae of Leguminosae are commonly used herbs in China, Japan, Korea, India and Thailand, with a long history of medicinal use. They are recorded in Chinese Pharmacopoeia, Japanese Pharmacopeia, Korea Pharmacopeia, Ayurveda Pharmacopoeia of India and Flora of Thailand. There are 15-20 species of Pueraria in the world, including 7 species and 2 varieties in China. At present, there are 6 species with medicinal value, such as Pueraria lobata and P. thomsonii. The main chemical components of the genus are isoflavones, flavonoids, terpenes, steroids, coumarins, puerarin glycosides and benzopyrans. A total of 240 compounds have been isolated and identified from this genus, and their pharmacological effects mainly include improvement of the cardiovascular system, antioxidant, hypoglycemic, antipyretic, anti-inflammatory, anti-alcoholic and estrogen-like effects. In this study, chemical constituents and pharmacological activities of Pueraria at home and abroad were systematically summarized, in order to provide references for the material basis, quality control and further development of Pueraria genus.

A reliable and rapid pharmacokinetic study of pueraria isoflavones using pueraria reference extractive substance in beagle plasma: Application to study of Yufeng Ningxin Tablets / 中草药·英文版

Objective: In order to evaluate the reliability and feasibility of pueraria reference extractive substance (RES) used in biological sample, the pharmacokinetics of 3′‑hydroxy puerarin (3′-HP), puerarin, 3′‑methoxy puerarin (3′-MP), and daidzein-8-C-apiosyl-(1-6)-glucoside (DAG) in beagle plasma following oral administration of Yufeng Ningxin Tablet were quantitated. Methods: A reliable and sensitive high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-QQQ-MS/MS) method developed with chromatographic separation was operated on a Merck C18 column, and acetonitrile-5 mmol/L ammonium was used as mobile phase in gradient elution. The plasma samples were deproteinized by acetone, detected by triple quadrupole mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. The pharmacokinetic parameters were calculated by Winnonlin 4.1. Results: The calibration curves of the reference extractive substance and standard substance methods were linear over the ranges 0.0417–11.3309 µg/mL and 0.0394–10.0000 µg/mL. The intra-day and inter-day precision of the two methods at three concentrations were less than 13.63%, and the average recoveries of 3′-HP, puerarin, 3′-MP, and DAG were more than 70.67%. The RSD of the mean plasma concentrations of the analytes calculated by the two methods was less than 5%, and cos (ϑ) = =1.000. Among the analytes, puerarin showed the highest blood concentration [(940 ± 185) ng/mL] and the longest retention time [(5 ± 1) h] in the dog's bodies. Conclusion: Pueraria reference extractive substance can be seen as an alternative to the standard substance to overcome the scarcity of standard substance for the analysis of biological samples.

Effects of pueraria total flavone on inflammatory cytokine and stat3 expression in brain tissue of exhaustive exercise rats / 中国组织工程研究

BACKGROUND: The physiological and pathological mechanisms underlying the inflammatory response of rat brain tissue caused by exhaustive exercise are very complicated. Studies have shown that Pueraria total flavonoids have anti-oxidative, neuroprotective, and cardio-cerebrovascular protective effects against brain injury. OBJECTIVE: To investigate the effect of Pueraria total flavone on inflammatory cytokines and signal transducer and activator of transcription-3 (STAT3) expression in rat brain tissue after exhaustive exercise. METHODS: Fifty healthy male Sprague-Dawley rats were randomly divided into a quiet control group, an exercise control group, low, middle, high dose Pueraria total flavone groups. Each training group conducted a 6-week sports training. At the end of 6-week exercise, the rats were confirmed to be exhausted. The rats were intragastrically administered 50, 100 and 200 mg/kg Pueraria total flavone in low, middle and high dose groups, respectively. Administration in each Pueraria total flavone group began at 30 minutes before exercise, once a day, and ended until the completion of the experiment. The activities of tumor necrosis factor-α, interleukin-8, interleukin-1β and interleukin-10 in serum and brain tissue of rats were determined by ELISA. The expression of STAT3 in rat brain tissue was detected by RT-PCR and western blot. The study protocol was approved by the Animal Ethics Committee of Guangxi Normal University (approval No. GXMU201703049). RESULTS AND CONCLUSION: The levels of tumor necrosis factor-α, interleukin-8, interleukin-1β and interleukin-10 in serum and brain tissue of exercise control rats were higher than those in the quiet control group (P < 0.01). The levels of tumor necrosis factor-α and interleukin-10 in serum and brain tissue of rats with middle and high dose of Pueraria total flavone were significantly lower than those in the exercise control group (P < 0.01). The levels of interleukin-8 and interleukin-1β in serum and brain tissue of rats with low, middle and high dose of Pueraria total flavone were significantly lower than those in the exercise control group (P < 0.01 or P < 0.05). The expression levels of STAT3 mRNA and protein in brain tissue of exercise control rats were significantly higher than those in the quiet control group (P < 0.01). The expression levels of STAT3 mRNA and protein in the brain tissue of rats with low, middle and high doses of Pueraria total flavone were significantly higher than those in the quiet control group (P < 0.01). To conclude, exhaustive exercise can cause inflammatory reaction and up-regulate STAT3 expression in rat brain tissue. The total flavonoids of Puerariae can regulate the expression of STAT3 in brain tissues and inhibit the inflammatory response of brain tissue, thus protecting damaged brain tissue.

Analysis of Flavonoids Chemical Components in Waste Material during Industrialization of Pueraria thomsonii Resources / 中国药房

OBJECTIVE：To study the composition and contents of flavonoids chemical components in waste material during industrialization of Pueraria thomsonii resources，and to provide reference for comprehensive development and reasonable utilization of the variety. METHODS ：Using“No. 2 Gange”of P. thomsonii from Jiangxi as objects ，UPLC-Q-TOF-MS and HPLC method were adopted to detect the components and contents of flavonoids in the root （with or without cortex ），cortex，flower， fibrous root ，stem，head and dregs （with or without cortex ）of P. thomsonii as well as dry matter of industrial wastewater （with or without cortex ）after precipitation of pueraria powder. RESULTS ：The linearity ，precision，repeatability，stability and recovery of the established method for content determination of 7 flavonoids（puerarin，daidzin，iridoxine-7-O-xylose glucoside ，genistin， iridin，daidzein and kakkalide ）were all in line with the requirements. Totally 12 kinds of flavonoids were identified ，among which the flavonoids in the root ，cortex，stem，fibrous root ，head and dregs of P. thomsonii as well as dry matter of industrial wastewater were the same ，mainly were puerarin ，daidzin，genistein，daidzein and malonyl-daidzein. The flower of P. thomsonii mainly included iridoxine- 7-O-xylose glucoside ，genistin，iridin，kakkalide，6″-O-xylosyldaidzein，but the components as puerarin ， daidzin and its aglycone were not be detected. The content of puerarin in the head of P. thomsonii was the highest （5.765％）. The contents of puerarin in root and dregs of P. thomsonii as well as dry matter of industrial waste-water in samples with cortex were all higher than in corresponding peeled sample. CONCLUSIONS ：The waste material from the industrialization of P. thomsonii resources contains a lot of flavonoids with rich species and high content ，and can be used as an important raw material for obtaining flavonoids such as puerarin.

Investigation of the estrogenic activity of Pueraria candollei variety mirifica extract on rats

Background: Pueraria candollei variety mirifica (PM) has been widely used as ingredient in many rejuvenating products. In this study, we aimed to assess the estrogenic activity of PM extract grown in Vietnam.Methods: Estrogenic activity of PM extract was estimated on immature female rats by using uterotrophic method to measure the weight of the reproductive organs. Estrogenic activity of PM extract also was investigated in mature female ovariectomized rats by evaluating the vaginal cells growth, reproductive organs weight, serum estradiol concentration.Results: Our results showed that PM extract at doses of 100 mg/kg, 200 mg/kg had increased the reproductive organs weight in immature rats and female ovariectomized rats. In addition, PM extract had increased the serum estradiol concentration and the vaginal cells growth by increasing the percentage of keratinocytes in female ovariectomized rats.Conclusions: Our results showed that PM extract has strong estrogenic activity in rats.

Analysis of genomic SSRs in Pueraria lobata and P. thomsonii and establishment of DNA identity card for different germplasms of P. thomsonii of Jiangxi province / 中国中药杂志

The type and frequency of simple sequence repeats( SSRs) in the genomes was investigated using the DNA sequence data of Pueraria lobata and P. thomsonii. Based on these SSRs,20 pairs of SSR primers were designed and 5 high polymorphism primer pairs were selected to analyze genetic diversity of 9 cultivars of P. thomsonii in Jiangxi province. The results showed that the 5 pairs of primers could generate 16 polymorphic alleles bands. The average polymorphism information content( PIC) of each SSR primer pair was 0. 600 7.According to the genetic similarity coefficients,the 9 cultivars of P. thomsonii can be classified into 6 germplasms. This study established DNA identity cards with 5 pairs of SSR primers for different germplasm resources of P. thomsonii in Jiangxi province,which provided reference information for the selection of fine germplasms of P. thomsonii and the theoretical basis for the study of Dao-di herbs.

Effect of bioadhesive excipients on absorption of total flavonids from Puerariae Lobatae Radix transporting across Caco-2 cell monolayer / 中草药·英文版

Objective: Pueraria total flavonids (PTF) can treat cardiovascular and cerebrovascular diseases, but it has poor membrane permeability and oral bioavailability. Some excipients, such as carbomer, chitosan, and hydroxypropyl methylcellulose, can improve the oral bioavailability. Traditional in vitro evaluation techniques, including the rat intestinal perfusion and cell line models, cannot evaluate PTF absorption and holistic transporters. Methods: This study evaluated excipients’ adhesiveness and effect on PTF transport across Caco-2 cell monolayer. cDNA microarrays identified gene expression changes in Caco-2 cells exposed to PTF and PTF with excipients, and revealed the mechanism underlying the effect of excipients on PTF absorption. Results: In vitro adhesion and transport experiments across Caco-2 showed that excipients had higher adhesiveness to gastric mucosa and transport efficiency across Caco-2 cells than PTF alone. The interaction of PTF with excipients significantly changed the expression of some genes, which might influence the absorption rate of PTF. Conclusion: Different bioadhesive polymers can improve intestinal absorption of PTF, which was related to some genes affiliated to the ATP-binding cassette (ABC) and solute carrier transporter (SLC) to some extent.

Applications of Pueraria lobata in treating diabetics and reducing alcohol drinking / 中草药·英文版

Pueraria lobata is one of the most important medicinal herbs used traditionally in China. According to Shanghan Lun (Treatise on Exogenous Febrile Disease), it has been used traditionally to relieve body heat, eye soring, dry mouth, headache associated with high blood pressure, and stiff neck problems. Modern studies in the 1970s revealed that isoflavonoids extracted from P. lobata were the bioactive components of an herbal remedy namely Yufeng Ningxin Tablets for the treatment of patients after stroke. This article reviews recent application of P. lobota in the treatment of diabetics and in reducing alcohol drinking. In view of its low toxicity profile, P. lobota stands an excellent chance to be developed as a phytomedicine for treating human diseases.

Clinical pharmacology and adverse reactions of Ge Tong Tong Luo Capsule in treating cerebral infarction / 中国药理学通报

The active ingredient of Ge Tong Tong Luo Capsule is the total flavonoids of Pueraria lobata, which mainly consists of puerarin, daidzein, daidzein-4V and other components. In the treatment of cerebral infarction, Ge Tong Tong Luo Capsule and its main pharmacological components have shown to inhibit the progression of atherosclerotic plaque, scavenge oxygen free radicals, anti-platelet aggregation, and inhibit neuronal apoptosis. In terms of adverse events, most of the studies were obvious adverse reactions caused by drugs. In rare studies, although there were adverse events such as palpitation and dry throat, they were relieved without treatment. This article introduces the pharmacological effects, clinical efficacy and adverse reactions of Ge Tong Tong Luo Capsule and its main pharmacological components, and provides a reference for clinical safe and rational drug use.

Network pharmacology-based study on mechanisms of antipyretic action of Pueraria radix / 药学学报

This study was designed to explore the antipyretic mechanism of Pueraria radix. The method of network pharmacology was used to determine the known ingredients corresponding to Pueraria radix, predict the drug-related gene /protein targets, and analyze the interplay between key ingredients and targets. Biological Information Annotation Databases (DAVID) was used to enrich the biological processes and pathways. The result of network analysis was validated by molecular docking. It was found that 49 active ingredients of Pueraria radix not only regulate 21 targets (e.g. PTGS2, EGFR), but also affect 11 biological processes (e.g. oxidation-reduction process, prostaglandin synthesis, positive regulation of fever generation and inflammatory response) and 7 metabolic pathways (arachidonic acid metabolism, serotonergic synapse and HIF-1, et al). Molecular docking results showed that more than 65% of the active ingredients could be well docked with key targets, and the relevant literatures indicated that the active components could inhibit the expression of PTGS2, which means the result has a high reliability. These results indicated that Pueraria radix may carry its pyretic action via a "multi-ingredients-multi-targets-multi-pathways" mode, which provides a scientific basis for further research and drug development.

Simultaneous Determination of 14 Kinds of Isoflavones in Pueraria radix by LC-MS/MS / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 14 kinds of isoflavones in Pueraria radix. METHODS:LC-MS/MS was adopted to detect 14 kinds of isaflavones in 14 batches of P. radix(P. radix:PL-1 to PL-7,P. thomsonii:PT-1 to PT-7). The determination was performed on Extend C18 with mobile phase consisted of methanol-water(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 22℃. The sample size was 5μL. Ion source was ESI source. The detection mode was negative ion detection. Scanning mode was MRM with jet voltage of -4500 V;ion source temperature was set at 600 ℃, and atomization gas was 60 psi. The auxiliary gas was 60 psi,collision gas was 7 psi,air curtain gas was 30 psi. SIMCA 13.0 software wasused for cluster analysis of above batches. RESULTS:The linear range of 14 kinds of isoflavones ranged 10-1000 ng/mL(puerarin of 10-5000 ng/mL,r>0.9900). RSDs of precision,stability and reproducibility tests were all lower than 5％. The recoveries were 95％-105％(RSD were 0.8％-4.5％,n=6). The total content of isoflavones were different significantly between P. radix and P. thomsonii. The contents of isoflavones in P. radix from different origins were different significantly. Among 14 kinds of isoflavones, the content of puerarin was the highest. Results of cluster analysis showed that 14 batches of P. radix could be clustered into 4 categories,including PL-2 as Ⅰ category,PL-3 and PL-4 as Ⅱ category,PL-5,PL-6 and PL-7 as Ⅲ category,other as Ⅳcategory. CONCLUSIONS:The method is simple and reproducible. It can be used for content determination of 14 kinds of isoflavones in P. radix and quality control.

Inhibitory Effect of Pueraria Isoflavones on Prostatic Hyperplasia in Rats / 中国药学杂志

OBJECTIVE: To investigate the inhibitory effect and possible mechanisms of puerariae isoflavone(PI) on prostatic hyperplasia induced by testosterone propionate.METHODS: Forty-eight male Wistar rats were randomly divided into six groups according to their body weight including normal control group, model group, 40, 80, 160 mg•kg-1•d-1 PI group, and finasteride positive control group. In addition to the sham operation for rats in the normal control group, the rats in other five groups performed castration surgery. After the restoration, the five groups of rats were subcutaneously injected with testosterone propionate (10 mg•kg-1•d-1) for 10 d to establish a benign prostatic hyperplasia model and then the subcutaneous injection was maintained every 2 d. High, middle and low dose PI groups were intragastrically administered (40, 80, 160 mg•kg-1•d-1) from the second day when the benign prostatic hyperplasia model was successfully constructed. The positive control group was given finasteride (1.0 mg•kg-1•d-1).Rats in normal and model groups were given an equal volume of saline for 28 d. After the last administration, the prostate and seminal vesicles were separated under anesthesia in rats, the wet weight and volume of the prostate and seminal vesicles were measured. The prostate and seminal vesicles index were calculated too. Rat blood was drawn and dihydrotestosterone(DHT) and estradiol (E2) in the serum were measured. Nitric oxide (NO), nitric oxide synthase (NOS), superoxide dismutase (SOD) and malondialdehyde (MDA) levels in prostate tissues were measured. The prostate tissue in each group was randomly selected for HE staining. The pathological structure of the prostate tissue was observed under an optical microscope.RESULTS: Compared with the normal control group, the prostate gland index and seminal vesicle gland index of the model group increased significantly (P<0.01), and the DHT and E2 levels in serum increased significantly (P<0.01). MDA content was increased while NO levels, NOS and SOD activities were significantly decreased (P<0.01). HE staining showed that the size of the prostate gland in the model group was different, there were obvious dilation, hyperplasia and papillary protrusions, and the cavity was full of pink and homogeneous density. The interstitial tissue showed obvious dilations of blood vessels, infiltration of inflammatory cells, and proliferation of fibrous connective tissues. Compared with the model group, the index and volume of prostate and seminal vesicles in the PI and positive control groups were significantly decreased (P<0.05 or P<0.01), and the levels of serum DHT and E2 in the middle and high doses PI groups were significantly lower (P<0.05 or P<0.01). In all treatment groups, MDA content was decreased and NO, NOS, and SOD levels were increased (P<0.05 or P<0.01) except the low-dose PI groups. There was moderately hyperplasia in low-dose PI group, mild prostatic hyperplasia in positive control group and middle-dose PI group, basically no hyperplasia in high-dose PI group.CONCLUSION: PI has a certain inhibitory effect on prostate hyperplasia induced by testosterone propionate, especially in the medium and high dose PI groups. The mechanism may be related to the effects of pueraria isoflavone on antioxidant,free radical scavenging in vivo, increasing NOS activity and increasing NO level.

Decoction of Isoflavone Content and Its Acid Hydrolysis Conversion Rate in the Flower of Pueraria lobata / 中国药房

OBJECTIVE:To establish a method for the decoction of isoflavone content and its acid hydrolysis conversion rate in the flower of Pueraria lobata. METHODS:Using the flowers of Pueraria lobata as raw material,the isoflavone with main com-ponent of tectoridin in the flower of P. lobata was prepared with ethanol,ethyl acetate extracted,ethanol recrystallized and puri-fied,and it was converted to tectorigenin with hydrolysis in hydrochloric acid. By screening the solvent and wavelength,UV spec-trophotometry was established to determine tectovidin and tectorigenin,and calculate the isoflavone content and acid hydrolysis con-version rate of tectoridin(expressed by the relative percentage of tectorigenin). It was compared with HPLC detection results,the accuracy of UV method was evaluated. RESULTS:The solvent was 70％ethanol solution containing 1％triethylamine,and the iso-flavone content was detected at wavelength of 339,274 nm. The linear range of tectoridin was 8.80-29.33 nmol/mL(r=0.9999). RSDs of precision(n=6),stability(n=5)and reproducibility(n=6)tests were lower than 1.94％;average recovery was 99.7％(RSD=1.77％,n=9). There were no statistical significances in the contents of total flavonoids (UV:17.64-25.55 nmol/mL vs. HPLC:17.39-24.40 nmol/mL) and the relative percentage of tectorigenin (UV:57.65％-87.59％ vs. HPLC:55.62％-91.14％). CONCLUSIONS:The established method is accurate,reliable,and can be used for the rapid determination of acid hydrolysis con-version rate of tectoridin.