基于网络药理学和基因表达数据库探讨银杏叶-葶苈子延缓心肌纤维化的作用机制

Ginkgo Biloba and Lepidii Semen Alleviate Myocardial Fibrosis Based on Network Pharmacology Combined with Gene Expression Omnibus Chip Analysis

目的:借助网络药理学分析,联合分子对接及基因表达芯片分析验证,探讨银杏叶-葶苈子延缓心肌纤维化的作用机制。方法:利用中药系统药理学数据库与分析平台筛选银杏叶-葶苈子的有效成分和靶点,通过人类基因数据库(GeneCards)、人类孟德尔遗传数据库(OMIM)获取心肌纤维化的靶点。将有效成分靶点与心肌纤维化靶点分别导入Cytoscape 3.8.0软件构建药物-成分-靶点网络、STRING平台构建蛋白-蛋白相互作用(PPI)网络,再利用Cytoscape自带的网络分析工具计算网络拓扑参数,分析获取关键靶点。将交集靶点通过R软件进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析。最后利用基因表达数据库(GEO)的芯片数据进行数据分析,并利用AutoDock软件进行有效成分与核心靶点分子对接,Pymol软件绘制分子对接模式图验证核心靶点。结果:通过相关数据库,获得药物有效成分39个,筛选出有效成分作用靶点116个,心肌纤维化靶点3322个。网络拓扑图分析获得转录因子P65(RELA)、雄性激素受体(AR)、前列腺素G/H合酶1(PTGS1)、肿瘤蛋白质P53(TP53)、核因子-κB(NF-κB)抑制剂α(NFKBIA)等关键靶点。GO分析主要涉及对化学应激的细胞反应、对类固醇激素反应、DNA结合转录因子结合、核受体活动、泛素蛋白连接酶等过程,KEGG富集分析主要有脂质和动脉粥样硬化、磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号通路、丝裂原活化蛋白激酶信号通路(MAPK)等信号通路。基因表达数据库分析验证表明,前6个核心靶点中,AR、RELA、TP53、NFKBIA、PTGS1差异有统计学意义(P<0.05)。分子对接结果显示主要活性成分与靶点均能自发结合且大部分结合能力良好。结论:银杏叶-葶苈子可能通过NF-κB、MAPK、PI3K/AKT信号通路等发挥抗炎、抗氧化应激作用,抑制成纤维细胞相关增殖和活化过程,减少细胞外基质沉积,从而发挥延缓心肌纤维化作用。

Objective:To explore the mechanism of the Chinese herb ginkgo biloba and lepidii semen alleviate myocardial fibrosis based on network pharmacology analysis,molecular docking and gene expression omnibus(GEO)chip analysis and verification.Methods:The pharmacologic database and analysis platform of traditional Chinese medicine system were used to screen the effective ingredients and targets of ginkgo biloba and lepidii semen.The target of myocardial fibrosis was obtained by GeneCards and online Mendelian inheritance in man(OMIM).The effective ingredients and myocardial fibrosis targets were introduced into Cytoscape 3.8.0 software to construct drug-ingredient-target network.The STRING platform builded protein-protein interaction(PPI)networks.The network analysis tools-Cytoscape was used to calculate network topology parameters,analyze and obtain key targets.The intersection targets were analyzed by R software for gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.The chip data of GEO was used for data analysis,and the molecular docking of effective ingredients and core targets was carried out by AutoDock software,and the molecular docking model diagram was drawn by Pymol software to verify the core targets.Results:Thirty nine effective ingredients were obtained,116 active ingredient action targets and 3322 myocardial fibrosis targets were screened.Key targets such as transcription factor P65(RELA),androgen receptor(AR),prostaglandin G/H synthase 1(PTGS1),tumor protein P53(TP53),and nuclear factor-κB(NF-κB)inhibitorα(NFKBIA)were identified by network topology analysis.GO enrichment analysis mainly involved cellular response to chemical stress,response to steroid hormones,DNA binding transcription factor binding,nuclear receptor activity,ubiquitin protein ligase,and other processes.KEGG enrichment analysis mainly included lipid and atherosclerosis,phosphatidylinositol 3-kinase(PI3K)/total protein kinase B(AKT)signaling pathway and mitogen-activated protein kinase(MAPK)signaling pathway,and other pathways.GEO gene chip analysis and verification showed that AR,RELA,TP53,NFKBIA,and PTGS1 were significantly different among the first 6 core targets(P<0.05).The results of molecular docking showed that the main effective ingredients and the target could spontaneously bind and most of the binding ability was better.Conclusion:Ginkgo biloba and lepidii semen shows anti-inflammatory and anti-oxidative stress effects through NF-κB,MAPK signaling pathway and PI3K/AKT signaling pathway,and other signaling pathways,by inhibiting fibroblast related proliferation and activation process,reducing extracellular matrix deposition,thereby alleviating myocardial fibrosis.