UPLC-Q-Exactive-MS/MS结合网络药理学分析五脉绿绒蒿的化学成分及其抗肝纤维化作用机制  

Chemical constituents and anti-liver fibrosis mechanism of Meconopsis quintuplinervia based on UPLC-Q-Exactive-MS/MS and network pharmacology

作　　者：孔苑琳 张剑光 苏宏娜 拉目加 兰建龙 杨正明 马权 黄艳菲 刘圆 KONG Yuan-lin;ZHANG Jian-guang;SU Hong-na;LA Mu-jia;LAN Jian-long;YANG Zheng-ming;MA Quan;HUANG Yan-fei;LIU Yuan(College of Pharmacy,Southuest Minzu University,Chengdu 610225,China;Qinzhou Provincial Healh School,Qinzhou 535000,China;Institute of Qinghai-Tibetan Plateau,Southuest Minzu University,Chengdu 610225,China;Sichuan Engineering laboratory of Qiang-Yi Medicinal Resources Protection and Uuilication Technology,Chengdu 610225,China;Qinghai-Tibetan Plateau Eithnic Medicinal Resources Protection and Utilication Key Laboratory of National Ethnic Affairs Comission of the People's Republic of China,Chengdu 610225,China)

机构地区：[1]西南民族大学药学院,四川成都610225 [2]四川省羌彝药用资源保护与利用技术工程实验室,广西钦州535000 [3]青藏高原民族药用资源保护与利用国家民委重点实验室,四川成都610225 [4]钦州市卫生学校,四川成都610225 [5]西南民族大学青藏高原研究院,四川成都610225

出　　处：《中国中药杂志》2022年第22期6097-6116,共20页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2019YFC1712305);四川省自然科学基金项目(2022NSFSC1605);2021年国家民委领军人才支持计划;中央高校基本科研业务费校级创新团队专项项目(ZYN2022067)。

摘　　要：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。In this study, UPLC-Q-Exactive-MS/MS was used to rapidly analyze the chemical constituents of Meconopsis quintupli-nervia, and the anti-liver fibrosis mechanism of M. quintuplinervia was preliminarily analyzed by network pharmacology, molecular docking, and cell experiments. The chemical constituents of M. quintuplinervia were identified according to the information of MS~1 and MS~2, as well as the data in the literature and databases. SwissTargetPrediction and TargetNet were used to predict the potential targets. The targets related to liver fibrosis were collected from GeneCards and OMIM. The protein-protein interaction(PPI) network was constructed by STRING. Cytoscape 3.6.1 was used to construct and analyze the "constituent-target-disease" network to obtain key targets and their corresponding constituents in the network. DAVID 6.8 was used for GO analysis and KEGG signaling pathway enrichment analysis. Finally, the preliminary verification was carried out by molecular docking and cell experiments. As a result, 106 chemical constituents were identified from M. quintuplinervia, including 66 flavonoids, 16 alkaloids, 18 phenolic acids, 1 anthocyanin, and 5 other constituents. Among them, 3 constituents were identified as potential new compounds, and 59 constituents were reported in M. quintuplinervia for the first time. Network pharmacology analysis showed that M. quintuplinervia presumably acted on AKT1, SRC, JUN, EGFR, STAT3, HSP90 AA1, MAPK3, and other core targets through luteolin, isorhamnetin, quercetin, apigenin, kaempferide, amurine, 2-methylflavinantine, allocryptopine, the multi and other active compounds, thereby regulating the PI3 K/AKT signaling pathway, pathways in cancer, proteoglycans in cancer, FoxO signaling pathway, and other pathways to exert anti-liver fibrosis effects. M. quintuplinervia extract(MQE) could significantly down-regulate PI3 K and AKT protein levels in the HSC-T6 cell model induced by TGF-β1, suggesting that MQE may have the ability to regulate the PI3 K/AKT signaling pathway.

关 键 词：五脉绿绒蒿 肝纤维化 UPLC-Q-Exactive-MS/MS 网络药理学 分子对接 

分 类 号：R284.1[医药卫生—中药学] R285[医药卫生—中医学]