基于网络药理学及分子对接技术探讨半夏泻心汤治疗胃食管反流病的作用机制

Study on the Action Mechanism of Pinellia Heart-Draining Decoction in Treating Gastroesophageal Reflux Disease Based on Network Pharmacology and Molecular Docking Technology

目的:基于网络药理学及分子对接技术探讨半夏泻心汤治疗胃食管反流病(gastroesophageal reflux disease,GERD)的作用机制。方法:基于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)检索半夏泻心汤组方中药活性成分,并通过Swiss Target Prediction数据库筛选活性成分相关作用靶点。在DisGeNET数据库、在线人类孟德尔遗传数据库(online mendelian inheritance in man,OMIM)和GeneCards数据库中检索疾病相关靶点及潜在作用靶点。将潜在作用靶点与其对应的活性成分导入Cytoscape 3.9.0软件构建“半夏泻心汤-活性成分-靶点”网络并进行拓扑分析筛选核心活性成分,构建蛋白质相互作用(protein-protein interaction,PPI)网络,拓扑分析筛选关键靶点,并构建关键靶点的PPI网络筛选核心靶点。将关键靶点导入DAVID数据库进行基因本体(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)信号通路富集分析。运用Autodock Vina软件将核心活性成分与核心靶点进行对接。结果:共获得140个中药活性成分、1111个相关靶点及4176个疾病相关靶点。将活性成分相关靶点与GERD靶点进行映射,得到412个半夏泻心汤治疗GERD的潜在作用靶点。“半夏泻心汤-活性成分-靶点”网络拓扑分析发现槲皮素、β-谷甾醇、黄芩素、山柰酚、豆甾醇、表小檗碱等是半夏泻心汤治疗GERD的核心活性成分。将潜在靶点导入STRING数据库构建PPI网络,拓扑分析得到42个关键靶点,关键靶点PPI网络分析发现,信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)、肿瘤蛋白p53(tumor protein p53,TP53)、热休克蛋白90AA1(heat shock proteins 90AA1,HSP90AA1)等为半夏泻心汤治疗GERD的核心靶点。GO功能富集分析共得到515个生物学过程条目、63个细胞组分条目及93个分子功能条目,KEGG信号通路富集分析筛选得到150条信号通路。分子对接显示,核心成分与核心靶点结合稳定。结论:半夏泻心汤中槲皮素、β-谷甾醇、豆甾醇等多种活性成分通过调控STAT3、TP53、HSP90AA1等多靶点参与肿瘤相关通路、人类巨细胞病毒感染等多条通路治疗GERD。

Objective:To study the action mechanism of Pinellia Heart-Draining Decoction on gastroesophageal reflux disease(GERD)based on network pharmacology and molecular docking technology.Methods:Based on the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP),the active ingredients of the Chinese medicinals of Pinellia Heart-Draining Decoction were retrieved,and the relevant action targets of the active ingredients were screened through the Swiss Target Prediction database.Disease-related targets were searched in DisGeNET database,online mendelian inheritance in man(OMIM)database and GeneCards database and disease targets,the intersection was the potential target of Pinellia Heart-Draining Decoction in treating GERD.The potential target and its corresponding active components were imported into the software of Cytoscape 3.9.0 to construct the network of"Pinellia Heart-Draining Decoction-active components-target",and the core active components were screened through topology analysis building protein-protein interaction(PPI)network was obtained;the key targets were screen through topology analysis.PPI network of key targets were constructed to screen core targets.The key targets were imported into DAVID database for gene ontology(GO)function enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)signal pathway enrichment analysis.Autodock Vina software was used to dock the core active ingredients with key targets.Results:There were 140 active components,1111 related targets and 4176 disease-related targets,OMIM database and GeneCards database.By mapping the relevant targets of active ingredients with GERD targets,412 potential targets of Pinellia Heart-Draining Decoction for GERD were obtained.The network topology analysis of"Pinellia Heart-Draining Decoction-active components-target"showed that quercetin,β-Glutosterol,baicalein,kaempferol,stigmasterol and epiberberine,etc.were the core active ingredients of Pinellia Heart-Draining Decoction in treating GERD.By importing potential targets into STRING database to build a PPI network,42 key targets were obtained through topology analysis.The analysis of key target PPI network showed that signal transducer and activator of transcription 3(STAT3),tumor protein p53(TP53)heat shock proteins 90 AA1(HSP90 AA1)etc.were the core targets of Pinellia Heart-Draining Decoction in treating GERD.GO function enrichment analysis obtained 515 biological process items,63 cell component items and 93 molecular function items,and KEGG signal pathway enrichment analysis screened 150 signal pathways.Molecular docking showed that the core components were stably bound to key targets.Conclusion:In Pinellia Heart-Draining Decoction,quercetin,β-Glutosterol,stigmasterol and other active ingredients participate in tumor-related pathways,human cytomegalovirus infection and other pathways to treat GERD by regulating multiple targets such as STAT3,TP53,HSP90 AA1,etc.