基于网络药理学和分子对接探讨逐风通痹汤治疗缺血性脑卒中的潜在作用靶点

The potential targets of Zhufeng Tongbi decoction in treating ischemic stroke based on network pharmacology and molecular docking

目的:通过网络药理学及分子对接探讨逐风通痹汤治疗缺血性脑卒中的潜在作用靶点。方法:利用中药系统药理学数据库与分析平台(TCMSP)与文献查询获得逐风通痹汤的有效成分及对应靶点。从Genecard、DrugBank、OMIM数据库获取缺血性脑卒中的疾病靶点,用Cytoscape 3.8.1构建逐风通痹汤-有效成分-靶点网络。在线使用Venny 2.1.0对逐风通痹汤靶点和缺血性脑卒中靶点取交集,利用STRING 11.5数据库和Cytoscape 3.8.1建立蛋白质的交互作用网络,以解析蛋白质-蛋白质相互作用(PPI)关系,并找出核心靶点。利用DAVID和微生信对交集靶点展开基因本体论(GO)功能和京都基因与基因组百科全书(KEGG)通路富集分析。采用PyMOL程序和AutodockTools 1.5.7软件对主要活性成分与核心靶点进行处理和分子对接,以评估分子之间的结合能力。结果:共挖掘到逐风通痹汤158个活性成分和120个潜在靶点;与1676个缺血性脑卒中相关靶点相互映射,找到交集靶点89个,核心成分主要包括油酸、槲皮素、豆甾醇、山柰酚、丹参酮等,PPI网络分析筛选出19个核心靶点,包括过氧化物酶体增殖物激活受体γ(Peroxisome Proliferator Activated Receptor Gamma,PPARG)、丝裂原活化蛋白激酶(Mitogen-Activated Protein Kinase,MAPK)3、前列腺素内过氧化物合酶2(Prostaglandin-Endoperoxide Synthase 2,PTGS2)、过氧化物酶体增殖物激活受体α(Peroxisome Proliferator-activated Receptor alpha,PPARA)、雌激素受体(Estrogen Receptor,ESR)1、血管紧张素转换酶(Angiotensin-converting Enzyme,ACE)等。推测这些靶点可能为逐风通痹汤治疗缺血性脑卒中的潜在靶点,GO富集分析分别富集在85个生物过程、87个细胞组分和89个分子功能,KEGG富集分析主要涉及5-羟色胺能突触、谷氨酸能突触、环磷酸腺苷(Cyclic Adenosine Monophosphate,cAMP)信号通路和过氧化物酶体增殖物激活受体(Peroxisome Proliferator Activated Receptor,PPAR)信号通路。分子对接结果显示,逐风通痹汤中的豆甾醇、丹参酮可作为主要活性物质,与缺血性脑卒中的核心靶点结合活性较强。结论:逐风通痹汤多靶点、多途径作用于缺血性脑卒中患者,为后续治疗缺血性脑卒中的分子机制研究提供方向。

Objective:To explore the potential targets of Zhufeng Tongbi decoction(逐风通痹汤)in the treatment of ischemic stroke through network pharmacology and molecular docking.Methods:The active components and corresponding targets of Zhufeng Tongbi decoction were obtained by means of systematic pharmacology database and analysis platform(TCMSP)and literature inquiry.Disease targets of ischemic stroke were obtained from Genecard,DrugBank and OMIM databases,and the Zhufeng Tongbi decoction active ingredient-target network was constructed by Cytoscape 3.8.1.Venny 2.1.0 was used online to take intersection between the target of Zhufeng Tongbi decoction and the target of ischemic stroke,and String 11.5 databases and Cytoscape 3.8.1 were used to establish the protein interaction network,so as to analyze the proteinprotein interaction(PPI)relationship and find out the core target.The gene ontology(GO)function and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed for intersection targets using DAVID and Michenshin.PyMOL program and AutodockTools 1.5.7 software were used for processing and molecular docking of main active ingredients with core targets to evaluate the binding ability between molecules.Results:A total of 158 active components and 120 potential targets of Zhufeng Tongbi decoction were discovered.It mapped with 1676 ischemic strokerelated targets and found 89 intersection targets,the core components mainly included oleic acid,quercetin,stigmasterol,kaempphenol,tanshinone,etc.Through PPI network analysis,19 core targets were selected,including PPARG,MAPK3,PTGS2,PPARA,ESR1,ACE,etc.It could be speculated that these targets may be potential targets for Zhufeng Tongbi decoction in the treatment of ischemic stroke.GO enrichment analysis was concentrated in 85 biological processes,87 cell components,and 89 molecular functions,respectively.KEGG enrichment analysis mainly involved 5-HT synapse,glutamergic synapse,cAMP signaling pathway,and PPAR signaling pathway.The molecular docking results showed that stigasterol and tanshinone in Zhufeng Tongbi decoction could be used as the main active substances,which had strong binding activity with the core target of ischemic stroke.Conclusion:The multi-target and multi-channel effects of Zhufeng Tongbi decoction on patients with ischemic stroke provide a research direction for the molecular mechanism of subsequent treatment of ischemic stroke.