基于网络药理学方法探讨芪归通络口服液治疗短暂性脑缺血发作的作用机制

Mechanism of Qigui Tongluo Oral Liquid in the Treatment of Transient Ischemic Attack Based on Network Pharmacology

目的:通过网络药理学方法,探讨芪归通络口服液治疗短暂性脑缺血发作(TIA)的作用机制。方法:运用中药系统药理学数据库与分析平台、中医药百科全书数据库,结合文献检索,获取芪归通络口服液中8味中药的化学成分及作用靶点基因,于GeneCards数据库、人类孟德尔遗传综合数据库中获取TIA疾病对应的靶标基因,交集后获得疾病-药物蛋白靶基因。利用Cytoscape软件构建“单味药-活性成分-作用靶点-疾病”相互作用网络,通过STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络,分别通过WebGestalt富集分析工具和DAVID数据库,对相关潜在作用靶点进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。利用Autodock软件对筛选出的蛋白与芪归通络口服液活性成分进行分子对接。结果:在芪归通络口服液中共筛选出87个与TIA相关的潜在活性成分和40个潜在作用靶点,主要活性成分包括蚓激酶、槲皮素、β-谷甾醇、木犀草素和异黄酮等,芪归通络口服液治疗TIA的关键靶点包括雌激素受体1、胱天蛋白酶3、凝血因子7、血管内皮生长因子A、纤溶酶原和P2Y嘌呤受体12等。GO功能富集分析显示,生物过程主要涉及多细胞生物过程、刺激反应和生物调节。KEGG通路富集分析得到77条代谢通路,主要涉及缺氧诱导因子1信号通路、血小板活化信号通路、Ras相关蛋白1信号通路和细胞流体切应力与动脉粥样硬化信号通路等。分子对接结果显示,筛选的靶点受体蛋白与活性成分具有较好的结合活性。结论:网络药理学研究结果揭示了芪归通络口服液治疗TIA的多成分、多靶点、多途径的作用特点,并预测了其可能的活性成分、作用通路和关键靶点,为其药效物质基础和作用机制研究提供了理论依据。

OBJECTIVE: To investigate the mechanism of Qigui Tongluo oral liquid in the treatment of transient ischemic attack(TIA) based on network pharmacology. METHODS: Chemical ingredients and target genes of 8 traditional Chinese medicines in Qigui Tongluo oral liquid were obtained by using Traditional Chinese medicine systems pharmacology database and analysis platform, encyclopedia of traditional Chinese medicine and literature search. Targets related to TIA were screened through GeneCards database and on-line Mendelian inheritance in man, the interactive targets of Qigui Tongluo oral liquid and TIA were further acquired. The interaction network of “single drug-active ingredient-target-disease” was constructed by Cytoscape software, and the protein-protein interaction(PPI) network was constructed by STRING database. WebGestalt enrichment analysis and DAVID database were used to conduct gene ontology(GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis for related potential targets. Autodock software was used to molecularly dock the selected proteins with the active ingredients of Qigui Tongluo oral liquid. RESULTS: A total of 87 potential active ingredients and 40 potential targets related to TIA were screened out from Qigui Tongluo oral liquid, the main active ingredients included lumbrokinase, quercetin, β-sitosterol, luteolin and isoflavone. Key targets of Qigui Tongluo oral liquid in the treatment of TIA included ESR1, CASP3, F7, vascular endothelial growth factor A, PLG and P2 RY12. GO enrichment analysis showed that biological processes mainly involved multicellular organismal process, response to stimulus and biological regulation. KEGG pathway enrichment analysis showed 77 metabolic pathways, mainly included hypoxia-inducible factor 1 signaling pathway, Platelet activation signaling pathway, Rap1 signaling pathway, Fluid shear stress and atherosclerosis signaling pathway, etc. Molecular docking showed that the bioactive compounds had good binding activity with the protein receptors of key target genes. CONCLUSIONS: Results of network pharmacology reveals the characteristics of Qigui Tongluo oral liquid in the treatment of TIA with multiple ingredients, multiple targets and multiple pathways, and predicts its possible active ingredients, action pathways and key targets, providing theoretical basis for the study of pharmacodynamic substance basis and mechanism of action.