为探讨菊茎叶总黄酮(total flavonoids from stems and leaves of Chrysanthemum morifolium,TFCSL)抗氧化应激的活性成分,阐明其药效物质基础和作用机制.采用HPLC建立不同批次TFCSL指纹图谱;以高浓度葡萄糖诱导人脐静脉内皮细胞建立氧化损伤模型,将丙二醛含量、乳酸脱氢酶含量和超氧化物歧化酶活性作为药效指标;采用灰色关联度和偏最小二乘法分析其谱-效关系确定抗氧化药效物质基础;基于网络药理学结合分子对接探究核心靶点及作用通路.从12批次TFCSL指纹图谱中确定12个共有峰,指认其中9个化学成分;各批次总黄酮样品均可减少细胞凋亡、降低丙二醛及乳酸脱氢酶含量、提高超氧化物歧化酶活性;综合2种数学模型确定峰5(芹菜素-7-O-β-D-葡萄糖苷)、峰6(异绿原酸C)、峰7(香叶木素-7-O-β-D-葡萄糖苷)为抗氧化物质基础;筛选出的3个活性成分作用于抗氧化应激的33个靶点;关键靶点为TNF、CASP3、EDNRA、XDH、PTGS2、MMP2,主要涉及脂质和动脉粥样硬化信号通路、IL-17信号通路、糖尿病并发症AGE-RAGE信号通路、TNF通路、MPKA通路等信号通路;分子对接结果显示活性成分与关键靶点之间均有较好的结合力.表明TFCSL抗氧化应激的物质基础可能为芹菜素-7-O-β-D-葡萄糖苷、异绿原酸C、香叶木素-7-O-β-D-葡萄糖苷,推测通过TNF、CASP3等靶点作用动脉粥样硬化信号通路、IL17信号通路发挥作用,体现菊茎叶多成分、多靶点抗氧化的作用特点.
Screening of antioxidative material basis of total flavonoids from stems and leaves of Chrysanthemum morifolium based on the spectrum-effect relationship and its mechanism
In order to explore the antioxidant chemical constituents of total flavonoids from stems and leaves of Chrysanthemum morifolium(TFCSL),and to reveal its pharmacodynamic material basis and mechanism of action.In this study,HPLC was used to establish the fingerprints of different batches of TFCSL;high glucose concentration induced oxidative damage model in human umbilical vein endothelial cells,and malondialdehyde,lactate dehydrogenase and superoxide dismutase activities were used as indicators of drug efficacy;gray correlation and partial least squares were used to analyze the spectral-efficacy rela-tionship to determine the antioxidant efficacy substances of TFCSL;based on network pharmacology combined with molecular docking to explore the core targets and pathways of action.Twelve common peaks were identified from the fingerprints of 12 batches of TFCSL,and 9 chemical components were designated;all batches of total flavonoid samples reduced apoptosis,low-ered malondialdehyde and lactate dehydrogenase content,and increased superoxide dismutase activity;peak 5(apigenin-7-O-β-D-glucoside),peak 6(isochlorogenic acid C)and peak 7(diosmetin-7-O-β-D-glucoside)were selected as the basis for TFCSL antioxidant substances by integrating two mathematical models;the three screened active ingredients acted on 33 tar-gets of anti-oxidative stress;the key targets were TNF,CASP3,EDNRA,XDH,PTGS2,MMP2,etc.,which were mainly in-volved in signaling pathways such as lipid and atherosclerosis signaling pathways,IL-17 signaling pathway,diabetic complica-tions AGE-RAGE signaling pathway,TNF pathway,MPKA pathway,and other signaling pathways;the molecular docking re-sults showed that there was good binding between the active ingredients and the key targets.It indicates that the material basis of TFCSL anti-oxidative stress may be apigenin-7-O-β-D-glucoside,isochlorogenic acid C,diosmetin-7-O-β-D-glucoside,and it is speculated that it plays a role in the role of atherosclerosis signaling pathway and IL17 signaling pathway through the targets of TNF and CASP3,reflecting the role of multi-components and multi-targets of stems and leaves of Chrysanthemum morifoli-um in the function of anti-oxidation.
stems and leaves of Chrysanthemum morifoliumtotal flavonoidsoxidative stressspectrum-effect relationship
万方数据
维普
