目的:基于网络药理学和分子对接技术探讨川牛膝治疗继发性闭经(secondary amenorrhea,SA)的作用机制.方法:从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)检索川牛膝的活性成分及作用靶点,从GeneCards数据库获取SA的疾病靶点,采用R软件得到两者的交集靶点.借助STRING数据库构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,通过Cytoscape 3.8.2软件进行聚类分析和拓扑分析从而筛选核心靶点.应用R软件进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析(P＜0.05).最后,采用Autodock Vina软件进行分子对接验证.结果:川牛膝活性成分4个,即槲皮素、β-谷甾醇、甜菜素、红苋甾酮;川牛膝作用靶点178个,SA相关靶点1 366个,两者的交集靶点56个;基于Net-work Analyzer插件的拓扑分析筛选得到9个核心靶点,包括丝氨酸/苏氨酸蛋白激酶(serine/threorine protein kinase,AKT1)、丝裂原活化蛋白激酶1(mitogen activated protein kinase 1,MAPK1)、雌激素受体 1(estrogen receptor 1,ESR1)等.GO 功能分析主要涉及对类固醇激素的反应、活性氧代谢过程、上皮细胞增殖等;KEGG通路135条,主要包括H1F-1信号通路、内分泌抵抗、雌激素信号通路等.分子对接显示,活性成分(槲皮素、β-谷甾醇)与核心靶点(AKT1、MAPK1)具有较好的结合活性.结论:川牛膝治疗SA具有多成分、多靶点、多途径的特点,其可能通过槲皮素、β-谷甾醇等成分作用于AKT1、MAPK1、ESR1等靶点,调控雌激素信号通路、内分泌抵抗等通路.
Study on the Mechanism of Action of Chuanniuxi in the Treatment of Secondary Amenorrhea Based on Network Pharmacology and Molecular Docking
Objective:To explore the mechanism of action of Chuanniuxi(Cyathulae Radix)in the treatment of secondary amenorrhea(SA)based on network pharmacology and molecular docking technology.Methods:The active ingredients and targets of Chuanniuxi were retrieved from the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP),and the disease targets of SA were obtained from the GeneCards database.The intersection targets of the two were obtained by R software.With the help of STRING database,the protein-protein interaction(PPI)network was constructed,and the cluster analysis and topology analysis were performed by Cytoscape 3.8.2 software to screen the core genes.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed by R software.Finally,the molecular docking verification was performed by AutoDock-Tools software.Results:A total of 4 active ingredients of Chuanniuxi were identified,namely quercetin,β-sitosterol,betaine,and rote-none.There were 97 targets of action,1 366 targets of SA,and 56 intersection targets of the two;and 9 core targets were screened out based on the topological analysis of Network Analyzer plug-in,including protein kinase B1(AKT1),mitogen activated protein kinase 1(MAPK1),estrogen receptor 1(ESR1),etc.GO functional analysis mainly involved the response to steroid hormones,reactive oxygen species metabolism,epithelial cell proliferation,etc.There were 135 KEGG pathways,mainly including HIF-1 signaling pathway,endo-crine resistance,estrogen signaling,etc.Molecular docking showed that the active ingredients(including quercetin and β-sitosterol)and the core targets(including AKT1 and MAPK1)had good binding activity.Conclusion:Chuanniuxi has the characteristics of multi-com-ponents,multi-targets,and multi-pathways in the treatment of SA.It may act on AKT1,MAPK1,ESR1 and other targets through querce-tin,β-sitosterol and other ingredients,and regulate endocrine resistance,estrogen signaling and other pathways.
万方数据
维普
