基于UPLC-Q-TOF-MS/MS和网络药理学方法探讨五味子藤茎保肝的潜在药效物质及作用机制  

Potential pharmacodynamic substances and mechanism of Schisandra chinensis rattan stems in liver protection based on UPLC-Q-TOF-MS/MS and network pharmacology

作　　者：曲中原 罗龙坦 张文君 胡扬 杨小龙 邴一凡 王淇漩 李文兰 邹翔[2,3] QU Zhong-yuan;LUO Long-tan;ZHANG Wen-jun;HU Yang;YANG Xiao-long;BING Yi-fan;WANG Qi-xuan;LI Wen-lan;ZOU Xiang(Teaching and Research Department of Chinese Pharmacy,College of Pharmacy,Harbin University of Commerce,Harbin 150076,China;Pharmaceutical Engineering Technology Research Center,College of Pharmacy,Harbin University of Commerce,Harbin 150076,China;Harbin Yuzhou Rentai Pharmaceutical Engineering Research Center Co.,Ltd.,Harbin 150076,China)

机构地区：[1]哈尔滨商业大学药学院中药学专业教研室,黑龙江哈尔滨150076 [2]哈尔滨商业大学药学院药物工程技术研究中心,黑龙江哈尔滨150076 [3]哈尔滨宇洲仁泰药物工程研究中心有限公司,黑龙江哈尔滨150076

出　　处：《中草药》2022年第8期2407-2416,共10页Chinese Traditional and Herbal Drugs

基　　金：黑龙江省重点研发计划指导类项目(GZ20210110);黑龙江省博士后科研启动金资助项目(LBH-Q16133);哈尔滨商业大学青年创新人才支持计划项目(2020CX11);哈尔滨市科技攻关计划项目(2012DB3BS057)。

摘　　要：目的基于高效液相色谱-四极杆飞行时间串联质谱法(UPLC-Q-TOF-MS/MS)分析五味子Schisandra chinensis藤茎的主要成分,并结合网络药理学和分子对接方法预测保肝药效物质和潜在的作用靶点及通路。方法根据UPLC-Q-TOF-MS/MS二级质谱裂解碎片信息,并结合文献数据,对五味子藤茎化学成分进行分析鉴定;通过中药系统药理学数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP),从鉴定获得的化学成分中筛选出口服生物利用度(oral bioavailability,OB)≥30%且符合类药五原则的活性成分;运用TCSMP数据库和SwissTargetPrediction数据库查找和预测五味子藤茎的成分靶点;通过在线人类孟德尔遗传数据库(Online Mendelian Inheritance in Man,OMIM)、GeneCards数据库和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)数据库筛选疾病靶点。采用String11.5数据库和Cytoscape 3.7.2软件构建PPI网络,并筛选核心靶点。利用DAVID 6.8进行基因本体(gene ontology,GO)注释和KEGG通路富集分析,并通过AutoDockTools 1.5.6软件对五味子藤茎的活性成分与作用靶点进行分子对接验证。构建酒精致肝细胞损伤体外模型,并考察五味子藤茎提取物对肝损伤细胞的保护作用。结果从五味子藤茎中共鉴定出40个主要成分,包括木脂素类、黄酮类、有机酸类化合物;进一步筛选出活性成分12个,活性成分与疾病交集靶点118个;通过网络拓扑筛选获得核心靶点16个;富集分析发现五味子藤茎保肝作用主要涉及的通路包括:肿瘤坏死因子(tumor necrosis factor,TNF)、丙型肝炎(hepatitis C)、乙型肝炎(hepatitis B)、血管内皮生长因子(vascular endothelial growth factor,VEGF)信号通路等。分子对接结果表明,五味子甲素、五味子乙素、五味子丙素、五味子酯甲、槲皮素、戈米辛M2、戈米辛R等成分可与表皮生长因子(epidermal growth factoObjective To analyze the main components of Schisandra chinensis rattan stems(SCRS)by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS),and predict the hepatoprotective substances and potential targets and signaling pathways by network pharmacology and molecular docking.Methods According to the fragmentation of secondary mass spectrometry and literature data,the chemical constituents of SCRS were analyzed and identified.The active components with OB≥30%in accordance with the five principles of drugs were screened through the TCMSP database.The potential targets of active components of SCRS were checked in TCMSP database and predicted by SwissTargetPrediction online platform.The“hepatitis”and“liver injury”were selected as key words to screen disease targets with OMIM,GeneCards and KEGG databases.PPI network was constructed using String11.5 database and Cytoscape 3.7.2 software,and core targets were screened.DAVID 6.8 database was used for gene ontology(GO)anotation and KEGG pathway enrichment analysis.AutoDockTools 1.5.6 software was used to verify the molecular docking between the active components and the key targets of SCRS.The in vitro model of liver cell injury was established and the protective effect of SCRS extract on liver cell injury was investigated.Results A total of 40 main components were identified in SCRS,including lignans,flavonoids,organic acids and so on.12 active ingredients were further screened,and 118 intersection targets of active ingredients and diseases were obtained.Sixteen core targets were obtained by network topology screening.Enrichment analysis showed that the main pathways involved in the hepatoprotective effect of SCRS included TNF,hepatitis C,hepatitis B and VEGF signaling pathways.The results of molecular docking showed that ingredients such as schisandrin A,schisandrin B,schisandrin C,schisantherin A,quercetin,gomixin M2 and gomixin R could interact with targets including EGFR,PIK3CA,TNF,Caspase-3 and other proteins

关 键 词：五味子藤茎 保肝 活性成分 网络药理学 分子对接 五味子甲素 五味子乙素 五味子丙素 五味子酯甲 槲皮素 戈米辛M2 戈米辛R 

分 类 号：R284.1[医药卫生—中药学] R285[医药卫生—中医学]