基于网络药理学和分子对接技术探讨柴胡-桂枝治疗急性缺血性脑卒中的作用机制

Mechanism of Radix Bupleuri-Cassia Twig in the Treatment of Acute Ischemic Stroke Based on Internet Pharmacology and Molecular Docking Technology

目的:应用网络药理学方法,探讨柴胡-桂枝治疗急性缺血性脑卒中的核心成分、关键靶点、生物学功能及信号通路,并通过分子对接技术初步验证。方法:通过中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)获取柴胡-桂枝活性成分,经Uniprot数据库转换为人源基因。通过人类孟德尔遗传综合数据库(online mendelian inheritance in man,OMIM)和人类基因组注释数据库(Gene Cards)获得急性缺血性脑卒中的疾病基因。筛选出药物与疾病的共同基因,借助Cytoscape软件制作"活性成分-疾病靶点"网络,分别通过STRING数据库和Metascape数据库对交集基因进行蛋白质-蛋白质相互作用分析和富集分析。借助Auto Dock软件进行分子对接验证,通过Py Mol软件绘制对接结果图。结果:共获得柴胡-桂枝抗急性缺血性脑卒中的19个活性成分、150个靶点。筛选出槲皮素、山柰酚为核心活性成分,AKT1、IL-6、VEGFA、TNF、TP53和CASP3为核心靶点基因。富集分析结果显示,柴胡-桂枝抗急性缺血性脑卒中的靶点显著富集于癌症通路、AGE-RAGE通路、丙型肝炎通路和HIF-1信号通路等通路。分子对接结果表明,柴胡-桂枝的核心活性成分与核心靶点之间均有较强的结合活性,槲皮素和TNF蛋白结合性最强。结论:柴胡-桂枝的活性成分通过影响炎症反应、氧化应激、血管新生和细胞凋亡等生物学过程,涉及多个靶点基因及信号通路,从而发挥治疗急性缺血性脑卒中的作用,具有多成分、多靶点和多途径的特点。

OBJECTIVE: To analyze the core components,key targets,biological functions and signal pathways of radix bupleuri-cassia twig in the treatment of acute ischemic stroke by using the network pharmacology method,and to verify the effect of the method by molecular docking technology. METHODS: The active components from radix bupleuri and Cassia twig were obtained by raditional Chinese medicine systems pharmacology database and analysis platform( TCMSP) and converted into human genes by Uniprot database. The disease genes of acute ischemic stroke were obtained by OMIM and Genecards database. The common genes of drugs and diseases were screened,the“active components-disease targets ” network was constructed by Cytoscape software,and protein interaction analysis and enrichment analysis of the intersecting genes were performed by STRING database and Metascape database,respectively. Molecular docking was verified by Auto Dock software and docking results were drawn by PyMol software.RESULTS: A total of 19 active components and 150 targets of radix bupleuri-cassia twig in the treatment of acute ischemic stroke were obtained. Quercetin and kaempferol were selected as the core active components,AKT1,IL-6,VEGFA,TNF,TP53 and CASP3 were the core target genes. Enrichment analysis showed that the anti-acute ischemic stroke targets of radix bupleuri and Cassia twig were significantly enriched in cancer pathway,AGE-RAGE pathway,hepatitis C pathway and HIF-1 signaling pathway. The results of molecular docking showed that there was strong binding activity between the core active components and the core targets of radix bupleuri-cassia twig,and quercetin had the strongest binding activity with TNF protein. CONCLUSIONS: The active components of radix bupleuri-cassia twig play a role in the treatment of acute ischemic stroke by affecting the biological processes such as inflammatory response,oxidative stress,angiogenesis and apoptosis,which is characterized by multiple components,multiple targets and multiple pathways.