Puerariae Radix protects against ulcerative colitis in mice by inhibiting NLRP3 inflammasome activation

Ulcerative colitis(UC)is a common inflammatory disease of the gastrointestinal tract.Traditional Chinese medicine(TCM)has long been used in Asia as a treatment for UC and Puerariae Radix(PR)is a reliable anti-diarrheal therapy.The aims of this study were to investigate the protective effect of PR using the dextran sulfate sodium salt(DSS)-induced UC model in mice and identify molecular mechanisms of PR action.The chemical constituents of PR via ultra-performance liquid chromatography/tandem mass spectrometry and identified potential PR and UC targets using a network pharmacology(NP)approach were obtained to guide mouse experiments.A total of 180 peaks were identified from PR including 48 flavonoids,46 organic acids,14 amino acids,8 phenols,8 carbohydrates,7 alkaloids,6 coumarins and 43 other constituents.NP results showed that caspase-1 was the most dysregulated of the core genes associated with UC.A PR dose of 0.136 mg/g administered to DSS treated mice reversed weight loss and decreased colon lengths found in UC mice.PR also alleviated intestinal mucosal shedding,inflammatory cell infiltration and mucin loss.PR treatment suppressed upregulation of NOD-like receptor protein 3(NLRP3),cysteinyl aspartate-specific proteases-1(caspase-1),apoptosis-associated speck-like(ASC)and gasdermin D(GSDMD)at both the protein and m RNA expression levels.The addition of a small molecule dual-specificity phosphatase inhibitor NSC 95397 inhibited the positive effects of PR.These results indicated that PR exerts a protective effect on DSS-induced colitis by inhibiting NLRP3 inflammasome activation in mice.

Effects of Radix Puerariae, Radix Rehmanniae and Their Compatibility on Blood Glucose and Blood Lipids in Mice

[Objectives]To explore the effects of the compatibility of Radix Puerariae and Radix Rehmanniae on blood glucose and blood lipids in diabetic mouses.[Methods]Diabetic mouse model was established.The body weight and fasting blood glucose of mice were measured after 7 and 14 d of administration,and the biochemical indicators of blood lipids(TC,HDL-C,and LDL-C)were detected after 14 d of administration.[Results]Compared with the Radix Puerariae group and Radix Rehmanniae group,the compatibility group(1:2)had the best hypoglycemic effect(P<0.05),and TC and LDL-C in the compatibility group(2:1)significantly decreased(P<0.05),while HDL-C in the compatibility group(1:1)significantly increased(P<0.05).[Conclusions]Radix Puerariae,Radix Rehmanniae and their combination can reduce the blood glucose of diabetic mice.The compatibility group(1:2)had a significant hypoglycemic effect(P<0.05),and LDL-C in the compatibility group(2:1)significantly declined,while HDL-C in the compatibility group(1:1)rose significantly.

Response surface optimization of microwave-assisted extraction for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii

Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.

Extraction and separation of total flavonoids from Flos Puerariae and its antioxidant activity

Objective To study the optimum extraction and separation process of total flavonoids in the flowers of Flos Puerariae and its antioxidative activity.Methods The total flavonoids were extracted with assistance of ultrasonic wave and the content was determined at 265 nm wavelength by Spectrophotometric method.Orthogonal experiment L9(34)was carried out to investigate the effects of concentration of solvent,the ratio of material to liquid,time length of extraction,and frequency of extraction on extraction results of total flavonoids from Flos Puerariae.The extracted solution was purified by petroleum ether and ethanol sequently.Under these conditions,the total flavonoids was eluted gradiently with mixed mobile phase of methanol-chloroform solution in the silica gel column system,and then determined by UV scanning and HPLC.Fenton reaction was used to produce and detect hydroxyl radicals(·OH),and pyrogallol system was used to produce and detect the superoxide radical anion(O2-·).Results The optimum conditions were as follows;using 40%(V/V)methanol as extractor with the ratio of material to liquid at 1∶30,and extracting for 2.5 h a time for 3 times.The extraction yield of total flavonoids was 13.6%.Six isoflavone compounds were isolated from the flowers of Flos Puerariae by the method of silica gel column chromatography.Antioxidative test results showed good performance of flavonoid scavenging capacity in both hydroxyl radical system and superoxide radical system and its IC50 was 7.65 μg·mL-1 and 0.18 mg·mL-1,respectively.Conclusions This study provided scientific basis for further development and utilization of Flos Puerariae and make preparation for later pharmacological research.

Qualitative and quantitative evaluation of Flos Puerariae by using chemical fingerprint in combination with chemometrics method

In order to better control the quality of Flos Puerariae(FP),qualitative and quantitative analyses were initially performed by using chemical fingerprint and chemometrics methods in this study.First,the fingerprint of FP was developed by HPLC and the chemical markers were screened out by similarity analysis(SA),hierarchical clustering analysis(HCA),principal components analysis(PCA),and orthogonal partial least squares discriminant analysis(OPLS-DA).Next,the chemical constituents in FP were profiled and identified by HPLC coupled to Fourier transform ion cyclotron resonance mass spectrometry(HPLCFT-ICR MS).Then,the characteristic constituents in FP were quantitatively analyzed by HPLC.As a result,31 common peaks were assigned in the fingerprint and 6 of them were considered as qualitative markers.A total of 35 chemical constituents were detected by HPLC-FT-ICR MS and 16 of them were unambiguously identified by comparing retention time,UV absorption wavelength,accurate mass,and MS/MS data with those of reference standards.Subsequently,the contents of glycitin,genistin,tectoridin,glycitein,genistein,and tectorigenin in 13 batches of FP were detected,ranging from 0.4438 to 11.06 mg/g,0.955 to 1.726 mg/g,9.81 to 57.22 mg/g,3.349 to 41.60 mg/g,0.3576 to 0.989 mg/g,and 2.126 to 9.99 mg/g,respectively.In conclusion,fingerprint analysis in combination with chemometrics methods could discover chemical markers for improving the quality control standard of FP.It is expected that the strategy applied in this study will be valuable for further quality control of other traditional Chinese medicines.

The Protective Effect of a Puerariae flos Extract (Thomsonide) against Ethanol-Induced Gastric Lesions in Rats

Objectives: Puerariae flos has popularly been used to treat alcoholic disorders. However, the effect of Puerariae flos on alcoholic disorders in the gastrointestinal system has not been identified. We investigated the protective effect of an extract of Puerariae flos against the murine gastric mucosa. Methods: Thomsonide, the extracts containing large amounts of isoflavonoid and triterpenoid saponin, was obtained fr om Puerriae flos via Diaion HP-20 column chromatography using water and 99.5% ethanol. It was investigated whether thomsonide, as well as geranylgeranylacetone (teprenone), a popular anti-ulcer agent developed in Japan, had a cytoprotective effect that might be related to endogenous prostaglandins, which played an important role in preventing gastric mucosal lesions. Results: Thomsonide and teprenone inhibited ethanol-induced gastric lesions. Furthermore, thomsonide increased the production of PGE2 and 6-ketoPGF1α, a stable metabolite of PGI2, in the gastric mucosa, and protective effects of thomsonide, as well as teprenone, against ethanol-induced gastric lesions were attenuated by pretreatment with indomethacin. Conclusions: These findings suggest that thomsonide, as well as teprenone, has the gastro protective effect which may be related to the cytoprotective activity of endogenous prostaglandins. The results of this study also suggest that the gastro protective effect of thomsonide may partially mitigate alcoholic disorders in the gastrointestinal tract, and support our pharmacological belief that Puerariae flos is useful for treatment of alcoholic disorders.

Antioxidative activities of the aqueous extracts of Puerariae and Notoginseng

AIM:To study the antioxidative activities of the aqueous extracts of Puerariae and Notoginseng in several models in vitro and their dose-effect relationships.

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

OBJECTIVE: To 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).METHODS: Human fetal scleral fibroblasts(HFSFs)were divided into 5 groups:(a) untreated controls;(b) cells treated with ELF-EMFs;(c) cells treated with ELF-EMFs and puerarin 0.1 μM;(d) cells treated with ELF-EMFs and puerarin 1 μM;(e) 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) m RNA, protein expression were measured by Real-Time polymerase chain reaction, Western blot analysis, respectively.RESULTS: Puerarin reduced the inhibition in cell proliferation, MMP-2 activity, m RNA, protein expression of HFSFs exposed to ELF-EMFs and enhanced the COL1A1 m RNA and protein expression.CONCLUSION: Puerarin 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.

Effect of bioadhesive excipients on absorption of total flavonids from Puerariae Lobatae Radix transporting across Caco-2 cell monolayer

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.

葛根总黄酮提取工艺优化及其抑菌与抗氧化活性研究

对葛根[Pueraria montana(Loureiro)Merrill]总黄酮的抑菌和抗氧化活性进行研究。采用单因素试验结合响应面法优化葛根的水提工艺,利用紫外分光光度法和高效液相色谱法对葛根总黄酮进行表征,并运用琼脂扩散法和以DPPH·、ABTS+·和总还原能力为指标评价葛根总黄酮的生物活性。结果表明,优化后的葛根总黄酮提取工艺为料液比(m∶V,g∶mL)1∶16,提取时间69 min,提取3次,总黄酮得率为7.690%。优化后的葛根总黄酮中检出9种常见黄酮成分,其含量总占比为91.81%;葛根总黄酮对11株供试菌有不同的抑菌活性,并首次发现其对粪链球菌(Enterococcus faecalis)和苏云金芽孢杆菌(Bacillus thuringiensis)有中度敏感作用,MIC分别为250、500 mg/mL,具有较强的清除能力和总还原能力,其中对DPPH·、ABTS~+·自由基的IC_(50)分别为0.398、0.189 mg/mL。

乙醇冷凝回流与超声波辅助提取葛根药材总黄酮

以云南省、广东省、广西壮族自治区不同地方葛根(Pueraria edulis)为材料,通过乙醇冷凝回流和超声波辅助提取法提取葛根药材中总黄酮,探究不同提取方式对不同地区葛根药材总黄酮含量的影响。结果表明,云南省葛根最佳提取时间均为20 min,广东省、广西壮族自治区葛根最佳提取时间均是30 min;冷凝回流提取法中葛根黄酮的得率相对较好;冷凝回流提取法中云南省产葛根总黄酮含量最高,平均含量为0.2557%,广东省葛根总黄酮平均含量最低为0.2108%。

Preparation and Performance of Pueraria lobata Root Powder/Polylactic Acid Composite Films

Petroleum-based materials,such as plastic,are characterized by adverse environmental pollution;as a result,researchers have sought alternative degradable plastics that are environmentally friendly,such as polylactic acid(PLA).PLA has shown great potential to replace petroleum-based plastics.In this study,seven different samples of unmodified Pueraria lobata root powder(PRP)with different contents(i.e.,0,5,10,15,20,25,and 30 wt%)and three different modified PRPs(i.e.,treated with NaOH,NaOH-KH-550,and Formic)were used to reinforce polylactic acid(PLA)via solution casting process.These prepared PRP/PLA composite films were characterized using SEM,FTIR,UV-visible spectra analysis,TG,DSC,weight loss measurement(wt%),and mechanical measurements.The results showed that the PRP modified with KH-550(PRPK)intensified the interaction in the interface region between the PRP and the PLA matrix,thus increasing the tensile strength(54.5 MPa),elongation at break(2.8%),and Young’s modulus(3310 MPa)of the PRPK/PLA biofilms.Contact angle measurement showed that the PRP treatments contributed to the hydrophobicity of films.The transparency of PRP-10/PLA film atλ_(800)was 11.09%,and its UVA and UVB transmittance were 3.28 and 1.16,respectively.After blending PLA with PRP,the PRP/PLA composite films exhibited excellent biodegradability.In summary,PRPK improved the mechanical properties of PLA and prevented the films from ultraviolet light,suggesting that PRPK-5/PLA film could be used as packaging materials.

Research progress of modern pharmacological effects of Pueraria lobata and its compound clinical application

As a traditional Chinese medicine, Pueraria lobata has been widely recorded in Chinese herbal works of the past dynasties. Modern studies have found that its main chemical components are isoflavones, triterpenoids, saponins, etc., which have antipyretic, anti⁃inflammatory, anti⁃infection, blood pressure, blood sugar, blood lipid, and pharmacological effects such as improving reproductive function. Clinically, it is used in the treatment of diabetic diseases, gastrointestinal diseases, cardiovascular diseases, etc. in the form of traditional Chinese medicine compound prescriptions. This reseach aims to summarize related articles of Pueraria lobata in the past 5 years, focusing on its pharmacological effects and clinical application of the compound, and provide references for its further development and utilization.

粉葛中微量元素含量的测定

[目的]测定粉葛样品中微量元素的含量。[方法]采用火焰原子吸收光谱法测定7种粉葛中Mg、Fe、Mn和Zn 4种微量元素的含量。[结果]粉葛样品富含Mg、Fe、Mn和Zn元素;不同产地不同品种粉葛的微量元素含量差异明显,即使同一产地不同品种粉葛的微量元素含量也有差异。[结论]该研究可为粉葛资源的进一步开发利用提供参考。

Effects of Ethanol Condensation Reflux Extraction and Ultrasound-assisted Extraction on the Content of Total Flavonoids of Pueraria edulis

With Pueraria edulis from different habitats in Yunnan Province,Guangdong Province and Guangxi Zhuang autonomous region as materials,the medicinal materials of P.edulis were extracted by ethanol condensation reflux extraction and ultrasound-assisted extraction,and the effects of different extraction methods the contents of total flavonoids in P.edulis from different areas were compared.The results showed that the best extraction time for P.edulis in Yunnan Province was 20 min,and that for Guangdong and Guangxi Zhuang Autonomous Region was 30 min.The yield of total flavonoids from P.edulis extracted by the condensation reflux extraction method was relatively better.The content of total flavonoids in P.edulis in Yunnan was the highest with an average content of 0.2557%,while the average content of total flavonoids in P.edulis in Guangdong was the lowest with an average content of 0.2108%.

葛根多糖提取条件的优化及其抗氧化活性的研究

采用乙醇回流法从葛根(Radix Puerariae)渣中提取葛根多糖,在单因素试验的基础上设计正交试验优化多糖提取工艺,并测定提取的葛根多糖的抗氧化活性。结果表明,优化的乙醇回流法提取葛根多糖的工艺条件为回流时间2.0 h、乙醇体积分数75%、料液比1∶40(m∶V,g/mL)、回流温度95℃,葛根多糖提取率为1.209%。提取的葛根多糖对O2-.和H2O2均有一定的清除作用,浓度为200 mg/L时,其对O2-.和H2O2的清除率分别为18.52%和13.68%。

Molecular mechanism study of Radix Pueraria for the treatment of glioma based on network pharmacology

Radix Pueraria(RP)has a long history of dual-use in medicine and food,and is well known as“Asian ginseng”.Recently,some studies about the effect of RP against glioma have been reported.However,little is elucidated about the molecular mechanism of interaction.This study used network pharmacology and molecular docking techniques to clarify the molecular mechanism of RP in the treatment of glioma.Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)was applied to screen potential active ingredients of RP and their targets.Targets for glioma were obtained from Genecards,OMIM,and Therapeutic Target Database(TTD).The topological analysis of cross-target and core target interaction was conducted using protein-protein interaction analysis(PPI).With the application of bioinformatics,GO and KEGG enrichment analyses were further carried out.Molecular docking was used then to validate the binding affinity between active components and key targets.This study successfully identified 6 potential active ingredients,namely beta-sitosterol,daidzein,formononetin,genistein,ononin,and puerarin.The hub targets were AKT1,TP53,VEGRA,EGFR,and MAPK3.These targets were mainly involved in biological processes,such as RNA polymerase II-specific DNA-binding transcription factor binding,and positive regulation of cell migration in the membrane raft,and were regulated by PI3K-Akt pathway.Further study showed that almost all the compounds had good binding abilities with above-mentioned targets.In short,this study systematically revealed the anti-glioma mechanism of RP to provide reference for the treatment of glioma.

超声波辅助提取葛根黄酮的工艺研究

采用超声波提取法,研究了乙醇浓度、固液比、超声波提取时间、超声波功率4个因素对葛根(Puerariae lobata(Willd.)Ohwi)黄酮提取率的影响,并采用L8(27)正交试验确定超声波提取葛根黄酮的最优工艺。结果表明:超声波作用时间与固液比对葛根黄酮得率有显著影响(P<0.05),并且其交互影响显著(P<0.05)。优化的葛根黄酮提取条件为:乙醇浓度70%、料液比1∶25、超声功率240W、超声时间40min。

超声微波协同萃取葛根中葛根素工艺的研究

[目的]探究利用超声微波协同萃取葛根中葛根素的提取工艺,以期为葛根素的工业化生产提供新思路。[方法]采用单因素试验和正交试验,以葛根素得率为指标,分别考察微波功率、提取时间、乙醇浓度、料液比及浸泡时间对葛根素提取的影响。[结果]在超声微波协同作用下,葛根素的最佳提取工艺为:微波功率为250W,提取时间为25s,乙醇浓度为50%,料液比为1:20,浸泡时间为30min。在此条件下,葛根素得率为0.6376%。[结论]超声微波协同萃取葛根素具有萃取效率高、节省时间、节省溶剂、节能、污染小等优点。

葛根产地、品种、采收、加工及其与药材品质关系研究进展

通过检索中国知网、万方数据知识服务平台、重庆维普等数据库,总结产地、品种、采收、加工对葛根药材品质影响的研究进展,为制定葛根药材药材种植、采收、产地加工等规范提供理论依据。