基于网络药理学探讨四君子汤治疗溃疡性结肠炎的作用机制及实验验证

Mechanism and experimental verification of Sijunzi Decoction in treatment of ulcerative colitis based on network pharmacology

基于网络药理学探讨四君子汤治疗溃疡性结肠炎(ulcerative colitis,UC)的作用机制。利用中药系统药理分析平台(TCMSP)提取四君子汤的活性成分及对应靶点,借助Uniprot数据库对靶点进行规范化,通过GeneCards数据库和Disgenet数据库获取UC的相关靶点,并利用R语言筛选药物与疾病的交集靶点;通过Cytoscape软件构建四君子汤"活性成分-疾病靶点"可视化调控网络,利用STRING数据库构建蛋白相互作用网络,并通过Bioconductor平台进行基因本体(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)通路富集分析,并采用动物实验验证部分靶点。经数据库分析,共获得四君子汤135个活性成分,预测作用靶点114个,与UC共同靶点80个,核心靶点蛋白包括白细胞介素6(interleukin-6,IL-6)、半胱氨酸蛋白酶3(caspase-3,CASP3)、血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、表皮生长因子受体(epidermal growth factor receptor,EGFR)等;GO功能富集分析涉及102个条目,主要影响转录因子活性、酶活性、受体活性及生物化学过程调控等过程;KEGG通路富集分析得到120条,涉及到人类巨细胞病毒感染、癌症、细胞凋亡、炎症等通路。小鼠实验证实四君子汤可下调靶点蛋白IL-6和caspase-3的表达,抑制肠上皮细胞凋亡。四君子汤治疗UC是多成分、多靶点、多通路相互作用的结果,经实验证实,可能通过调控IL-6和caspase-3的表达,从而参与细胞凋亡、炎症等通路发挥效用。

To explore the mechanism of Sijunzi Decoction in the treatment of ulcerative colitis(UC) based on network pharmacology. The active components and corresponding targets of Sijunzi Decoction were extracted with Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets were standardized with the help of Uniprot database. The related targets of UC were obtained through GeneCards database and Disgenet database, and the intersection targets of drugs and diseases were screened by R language. The visual regulation network of "active ingredient-disease target" of Sijunzi Decoction was constructed by Cytoscape software, and the protein-protein interaction network was constructed by STRING database. The functional enrichment analysis of gene ontology(GO) and the enrichment analysis of Kyoto encyclopedia of genes and genomes(KEGG) pathway were carried out on Bioconductor platform, and some of the targets were verified by animal experiments. Through database analysis, a total of 135 active components of Sijunzi Decoction, 114 predicted targets and 80 common targets with UC were obtained. The core target proteins included interleukin 6(IL-6), caspase-3(CASP3), vascular endothelial growth factor A(VEGFA), epidermal growth factor receptor(EGFR) and so on. GO functional enrichment analysis involved 102 items, which mainly affected transcription factor activity, enzyme activity, receptor activity and biochemical process regulation. KEGG pathway enrichment analysis showed that 120 items were involved in human cytomegalovirus infection, cancer, apoptosis, inflammation and other pathways. Mouse experiments showed that Sijunzi Decoction could down-regulate the expression of target proteins IL-6 and caspase-3 and inhibit intestinal epithelial cell apoptosis. The treatment of UC with Sijunzi Decoction is the result of the interaction among multi-components, multi-targets and multi-pathways. It is proved by experiments that Sijunzi Decoction may play an effective role by regulating the expression of IL-6 and caspase-3, and getting involved in apoptosis, inflammation and other pathways.