利用UPLC-Q-Exactive-MS/MS技术快速分析五脉绿绒蒿的化学成分,通过网络药理学、分子对接技术和细胞实验初步分析五脉绿绒蒿抗肝纤维化作用机制。根据一级质谱和二级质谱信息,结合文献和数据库数据,对五脉绿绒蒿中的化学成分进行指认。...利用UPLC-Q-Exactive-MS/MS技术快速分析五脉绿绒蒿的化学成分,通过网络药理学、分子对接技术和细胞实验初步分析五脉绿绒蒿抗肝纤维化作用机制。根据一级质谱和二级质谱信息,结合文献和数据库数据,对五脉绿绒蒿中的化学成分进行指认。采用SwissTargetPrediction及TargetNet数据库预测成分潜在作用靶点,GeneCards、OMIM数据库获取肝纤维化相关靶点,STRING数据库构建靶点蛋白相互作用PPI网络,Cytoscape 3.6.1软件进行“成分-疾病-靶点”的构建与分析,获取网络中的关键靶点与其对应成分,DAVID 6.8数据库进行GO分析和KEGG信号通路富集分析,利用分子对接技术及细胞实验进行初步验证。结果显示从五脉绿绒蒿中指认了106个化合物,包括黄酮类66个,生物碱类16个,酚酸类18个,花青素类1个以及其他类成分5个,其中可能的新化合物3个,五脉绿绒蒿中首次指认的化合物59个;网络药理学分析表明,五脉绿绒蒿可能通过木犀草素、异鼠李素、槲皮素、芹菜素、山柰酚-4′-甲醚、黑水罂粟碱、甲氧基淡黄巴豆亭碱、别隐品碱等成分,作用于AKT1、SRC、JUN、EGFR、STAT3、HSP90AA1、MAPK3等靶点,调节PI3K/AKT、pathways in cancer、proteoglycans in cancer、FoxO等信号通路发挥抗肝纤维化作用。五脉绿绒蒿提取物(MQE)可显著下调TGF-β1诱导的HSC-T6细胞模型中PI3K和AKT蛋白水平,表明MQE可能具有调节PI3K/AKT信号通路的能力。研究结果说明五脉绿绒蒿抗肝纤维化是多成分、多靶点、多途径的作用特点,为五脉绿绒蒿进一步开展治疗肝病的物质基础、作用机制和质量标志物研究提供科学依据。展开更多
In the present study, we developed a rapid, sensitive and efficient ultra high-performance liquid chromatography-tandem mass spectrometric(UPLC-Q-Exactive-MS) method, and six bioactive constituents in San-Chen-pill(SC...In the present study, we developed a rapid, sensitive and efficient ultra high-performance liquid chromatography-tandem mass spectrometric(UPLC-Q-Exactive-MS) method, and six bioactive constituents in San-Chen-pill(SCP), including cholic acid(CA), hyodeoxycholic acid(HDCA), hydroxysafflor yellow A(HSYA), kaempferol-3-O-rutinoside(KFR), anhydrosafflor yellow B(AHSYB) and syringin(SRG), in rat plasma after oral administration of San-Chen-pill, were simultaneously determined. Plasma samples were pretreated with methanol for protein precipitation. Chromatography separation was performed on a SHIMADZU of shim-pack GIST C18 column using a gradient mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution. A tandem mass spectrometric detection with an electrospray ion source(HESI) interface was conducted in negative ionization mode. For all the six analytes of interest, the calibration curves were linear within the concentration rage of 0.50–1280 ng/m L with r≥0.99. The intra-day and inter-day precisions(in terms of relative standard deviation, RSD) were all below 12.7% for all six analytes. Extraction recovery, matrix effect and stability data all met the acceptance criteria of FDA guideline for bioanalytical method validation. In addition, San-Chen-pill is a traditional Mongolian herbal medicine that has been prepared with three medicinal herbs, Calculus bovis, Carthamus tinctorius L. and Concretio silicea bambusae, and it shows potent anti-anginal activity in all preparations with in a ratio of 1:1:1. The pharmacokinetic parameters of San-Chen-pill were reported herein for the first time, and this developed method was successfully used in a pharmacokinetic study for San-Chen-pill.展开更多
Objective:To rapidly identify the two morphologies and chemical properties of similar herbal medicines,Blumea riparia and B.megacephala as the basis for chemical constituent analysis.Methods:UPLC-Q-Exactive-MS/MS was ...Objective:To rapidly identify the two morphologies and chemical properties of similar herbal medicines,Blumea riparia and B.megacephala as the basis for chemical constituent analysis.Methods:UPLC-Q-Exactive-MS/MS was utilized for profiling and identification of the constituents in B.riparia and B.megacephala.Chemical pattern recognition(CPR)was further used to compare and distinguish the two herbs and to identify their potential characteristic markers.Then,an HPLC method was established for quality evaluation.Results:A total of 93 constituents are identified,including 54 phenolic acids,35 flavonoids,two saccharides,one phenolic acid glycoside,and one other constituent,of which 67 were identified in B.riparia and B.megacephala for the first time.CPR indicates that B.riparia and B.megacephala samples can be distinguished from each other based on the LC–MS data.The isochlorogenic acid A to cryptochlorogenic acid peak area ratio calculated from the HPLC chromatograms was proposed as a differentiation index for distinguishing and quality control of B.riparia and B.megacephala.Conclusion:This study demonstrates significant differences between B.riparia and B.megacephala in terms of chemical composition.The results provide a rapid and simple strategy for the comparison and evaluation of the quality of B.riparia and B.megacephala.展开更多
Background:Simiaowan(SMW),a well-known traditional Chinese medicine,has been employed to treat hyperuricemia(HUA)and gout for centuries.However,the bioactive components and underlying mechanisms have not been elucidat...Background:Simiaowan(SMW),a well-known traditional Chinese medicine,has been employed to treat hyperuricemia(HUA)and gout for centuries.However,the bioactive components and underlying mechanisms have not been elucidated.The objective of this study was to identify the active components and potential mechanisms of SMW by integrating pharmacological experimentation,serum pharmacochemistry,network pharmacology and molecular docking.Methods:HUA rats modelling by high-fat/high-sugar diet and potassium oxonate/adenine oral administration were used to evaluate the pharmacodynamic effects of SMW.UPLC-Q-Exactive-MS/MS was employed to detect the bioactive components present in SMW-containing serum.Network pharmacology and molecular docking were utilized to elucidate the potential targets and underlying mechanisms.Results:SMW effectively ameliorated HUA rats via the inhibition of uric acid(UA)production,promotion of UA excretion,improvement of lipid and glucose metabolic abnormalities,antioxidant,anti-inflammatory and anti-insulin resistance effects.A total of 73 compounds detected in SMW-containing serum were identified as potential active components,with alkaloids,flavonoids,organic acids,and terpenoids emerging as the primary active ingredients.Totally 203 corresponding targets were obtained as SMW anti-HUA/gout targets,which mainly participated in apoptosis,insulin resistance,TNF,PI3K-Akt,HIF-1,NF-κB,MAPK,IL-17 and TLR signaling pathways.Molecular docking indicated that active compounds(e.g.berberine,phellodendrine,quercetin,formononetin,ferulic acid)had superior binding abilities to the key targets(e.g.solute carrier family 22 member 12(URAT1),solute carrier family 22 member 6(OAT1),ATP-binding cassette sub-family G member 2(ABCG2),solute carrier family 2,facilitated glucose transporter member 9(GLUT9),xanthine dehydrogenase/oxidase(XDH),transcription factor p65(RELA),toll-like receptor 4(TLR4),prostaglandin G/H synthase 2(PTGS2),caspase-3(CASP3),insulin(INS)).Conclusion:SMW exerted regulatory influence over the disease network of HUA and gout through a multiplicity of components,targets,and pathways.Alkaloids,flavonoids,organic acids,and terpenoids were the primary active components,exerting anti-HUA/gout effects via antioxidant,anti-inflammatory,anti-insulin resistance,anti-apoptosis,inhibition of UA production,and promotion of UA excretion.This study revealed the active components and molecular mechanisms of SMW,providing insights into the development of natural products derived from SMW.展开更多
文摘利用UPLC-Q-Exactive-MS/MS技术快速分析五脉绿绒蒿的化学成分,通过网络药理学、分子对接技术和细胞实验初步分析五脉绿绒蒿抗肝纤维化作用机制。根据一级质谱和二级质谱信息,结合文献和数据库数据,对五脉绿绒蒿中的化学成分进行指认。采用SwissTargetPrediction及TargetNet数据库预测成分潜在作用靶点,GeneCards、OMIM数据库获取肝纤维化相关靶点,STRING数据库构建靶点蛋白相互作用PPI网络,Cytoscape 3.6.1软件进行“成分-疾病-靶点”的构建与分析,获取网络中的关键靶点与其对应成分,DAVID 6.8数据库进行GO分析和KEGG信号通路富集分析,利用分子对接技术及细胞实验进行初步验证。结果显示从五脉绿绒蒿中指认了106个化合物,包括黄酮类66个,生物碱类16个,酚酸类18个,花青素类1个以及其他类成分5个,其中可能的新化合物3个,五脉绿绒蒿中首次指认的化合物59个;网络药理学分析表明,五脉绿绒蒿可能通过木犀草素、异鼠李素、槲皮素、芹菜素、山柰酚-4′-甲醚、黑水罂粟碱、甲氧基淡黄巴豆亭碱、别隐品碱等成分,作用于AKT1、SRC、JUN、EGFR、STAT3、HSP90AA1、MAPK3等靶点,调节PI3K/AKT、pathways in cancer、proteoglycans in cancer、FoxO等信号通路发挥抗肝纤维化作用。五脉绿绒蒿提取物(MQE)可显著下调TGF-β1诱导的HSC-T6细胞模型中PI3K和AKT蛋白水平,表明MQE可能具有调节PI3K/AKT信号通路的能力。研究结果说明五脉绿绒蒿抗肝纤维化是多成分、多靶点、多途径的作用特点,为五脉绿绒蒿进一步开展治疗肝病的物质基础、作用机制和质量标志物研究提供科学依据。
基金Natural Science Foundation of Inner Mongolia(Grant No.2017BS0806)Science Fund of Inner Mongolia Education Department(Grant No.NJZZ19089)
文摘In the present study, we developed a rapid, sensitive and efficient ultra high-performance liquid chromatography-tandem mass spectrometric(UPLC-Q-Exactive-MS) method, and six bioactive constituents in San-Chen-pill(SCP), including cholic acid(CA), hyodeoxycholic acid(HDCA), hydroxysafflor yellow A(HSYA), kaempferol-3-O-rutinoside(KFR), anhydrosafflor yellow B(AHSYB) and syringin(SRG), in rat plasma after oral administration of San-Chen-pill, were simultaneously determined. Plasma samples were pretreated with methanol for protein precipitation. Chromatography separation was performed on a SHIMADZU of shim-pack GIST C18 column using a gradient mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution. A tandem mass spectrometric detection with an electrospray ion source(HESI) interface was conducted in negative ionization mode. For all the six analytes of interest, the calibration curves were linear within the concentration rage of 0.50–1280 ng/m L with r≥0.99. The intra-day and inter-day precisions(in terms of relative standard deviation, RSD) were all below 12.7% for all six analytes. Extraction recovery, matrix effect and stability data all met the acceptance criteria of FDA guideline for bioanalytical method validation. In addition, San-Chen-pill is a traditional Mongolian herbal medicine that has been prepared with three medicinal herbs, Calculus bovis, Carthamus tinctorius L. and Concretio silicea bambusae, and it shows potent anti-anginal activity in all preparations with in a ratio of 1:1:1. The pharmacokinetic parameters of San-Chen-pill were reported herein for the first time, and this developed method was successfully used in a pharmacokinetic study for San-Chen-pill.
基金supported by National Key Research and Development Program of China(No.2018YFC1706101)The Sichuan Key Research and Development Program(No.2021YFS0043)+1 种基金Leading Talent Support Program of National Civil Commission(2021)Special Fund of Natural Science for the Central Universities(No.2020NQN42).
文摘Objective:To rapidly identify the two morphologies and chemical properties of similar herbal medicines,Blumea riparia and B.megacephala as the basis for chemical constituent analysis.Methods:UPLC-Q-Exactive-MS/MS was utilized for profiling and identification of the constituents in B.riparia and B.megacephala.Chemical pattern recognition(CPR)was further used to compare and distinguish the two herbs and to identify their potential characteristic markers.Then,an HPLC method was established for quality evaluation.Results:A total of 93 constituents are identified,including 54 phenolic acids,35 flavonoids,two saccharides,one phenolic acid glycoside,and one other constituent,of which 67 were identified in B.riparia and B.megacephala for the first time.CPR indicates that B.riparia and B.megacephala samples can be distinguished from each other based on the LC–MS data.The isochlorogenic acid A to cryptochlorogenic acid peak area ratio calculated from the HPLC chromatograms was proposed as a differentiation index for distinguishing and quality control of B.riparia and B.megacephala.Conclusion:This study demonstrates significant differences between B.riparia and B.megacephala in terms of chemical composition.The results provide a rapid and simple strategy for the comparison and evaluation of the quality of B.riparia and B.megacephala.
基金supported by Natural Science Foundation of Guangdong Province(2021A1515010978 and 2021A1515012474)Basic research project of Shenzhen Science and Innovation Commission(JCYJ20210324121610029)Guangdong Provincial Key Areas Research and Development Program project Lingnan TCM Modernization(2020B1111120003).
文摘Background:Simiaowan(SMW),a well-known traditional Chinese medicine,has been employed to treat hyperuricemia(HUA)and gout for centuries.However,the bioactive components and underlying mechanisms have not been elucidated.The objective of this study was to identify the active components and potential mechanisms of SMW by integrating pharmacological experimentation,serum pharmacochemistry,network pharmacology and molecular docking.Methods:HUA rats modelling by high-fat/high-sugar diet and potassium oxonate/adenine oral administration were used to evaluate the pharmacodynamic effects of SMW.UPLC-Q-Exactive-MS/MS was employed to detect the bioactive components present in SMW-containing serum.Network pharmacology and molecular docking were utilized to elucidate the potential targets and underlying mechanisms.Results:SMW effectively ameliorated HUA rats via the inhibition of uric acid(UA)production,promotion of UA excretion,improvement of lipid and glucose metabolic abnormalities,antioxidant,anti-inflammatory and anti-insulin resistance effects.A total of 73 compounds detected in SMW-containing serum were identified as potential active components,with alkaloids,flavonoids,organic acids,and terpenoids emerging as the primary active ingredients.Totally 203 corresponding targets were obtained as SMW anti-HUA/gout targets,which mainly participated in apoptosis,insulin resistance,TNF,PI3K-Akt,HIF-1,NF-κB,MAPK,IL-17 and TLR signaling pathways.Molecular docking indicated that active compounds(e.g.berberine,phellodendrine,quercetin,formononetin,ferulic acid)had superior binding abilities to the key targets(e.g.solute carrier family 22 member 12(URAT1),solute carrier family 22 member 6(OAT1),ATP-binding cassette sub-family G member 2(ABCG2),solute carrier family 2,facilitated glucose transporter member 9(GLUT9),xanthine dehydrogenase/oxidase(XDH),transcription factor p65(RELA),toll-like receptor 4(TLR4),prostaglandin G/H synthase 2(PTGS2),caspase-3(CASP3),insulin(INS)).Conclusion:SMW exerted regulatory influence over the disease network of HUA and gout through a multiplicity of components,targets,and pathways.Alkaloids,flavonoids,organic acids,and terpenoids were the primary active components,exerting anti-HUA/gout effects via antioxidant,anti-inflammatory,anti-insulin resistance,anti-apoptosis,inhibition of UA production,and promotion of UA excretion.This study revealed the active components and molecular mechanisms of SMW,providing insights into the development of natural products derived from SMW.
