基于网络药理学探讨四神煎治疗类风湿关节炎的作用机制

目的:基于网络药理学探讨四神煎治疗类风湿关节炎(rheumatoid arthritis, RA)的作用机制。方法:通过中药系统药理数据库和分析平台(traditional chinese medicine systems pharmacology database and analysis platform, TCMSP)、中医药百科全书数据库(encyclopedia of traditional chinese medicine, ETCM)检索四神煎组方中药的主要化学成分,并结合文献进行筛选、补充;通过Pubchem有机小分子生物活性数据库检索化学成分的SMILES结构式及2D结构,经Swiss ADME数据库筛选后将符合标准的化学成分上传至Swiss Target Prediction数据库预测药物活性靶点;通过人类基因数据库(Gencards)、在线人类孟德尔遗传数据库(online mendelian inheritance in man, OMIM)、治疗靶标数据库(therapeutic target database, TTD)、DrugBank数据库获取RA疾病相关靶点;绘制韦恩图并获取四神煎活性成分与RA的共同靶点;基于String平台构建药物疾病共同靶点的蛋白互作网络;基于Metascape平台进行基因本体(gene ontology, GO)生物功能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;采用Cytoscape 3.7.1软件构建"成分-靶点-通路"网络,应用网络分析仪(network analyzer)功能获得各成分、靶点的网络拓扑参数,筛选四神煎发挥治疗RA作用的重要活性成分及核心靶点。结果:通过ETCM、TCMSP数据库结合文献检索获得60个四神煎活性成分及353个相关靶点;通过GeneCards、OMIM、TTD、DrugBank四个数据库获得1 277个RA潜在靶点;将药物成分作用靶点与RA相关靶点映射后,得到四神煎治疗RA的靶点151个;GO生物功能富集分析得到2 322个条目,其中生物过程2 089条,细胞组分79条,分子功能154条;KEGG通路富集分析获得174个信号通路,主要涉及白细胞介素-17(interleukin-17,IL-17)信号通路、肿瘤坏死因子(tumor necrosis factor, TNF)信号通路、Th17细胞分化、破骨细胞分化、核因子-κB(nuclear factor-kappa B,NF-κB)信号通路等;网络拓扑分析得出四神煎发挥治疗RA作用的重要活性成分有槲皮素、木犀草素、山柰酚、汉黄芩素等化合物,重要靶点基因主要有PTGS2、PTGS1、RELA、TNF、NFKB1、IL-6等。结论:四神煎可能通过槲皮素、木犀草素、山柰酚、汉黄芩素等活性成分调控PTGS2、PTGS1、RELA、TNF、NFKB1、IL-6等靶点基因的表达,进而调控IL-17信号通路、TNF信号通路、Th17细胞分化、破骨细胞分化等生物学通路发挥治疗RA的作用。

Study on the Mechanism of Sishen Decoction in the Treatment of Rheumatoid Arthritis Based on Network Pharmacology

Objective: To explore the mechanism of Sishen Decoction in the treatment of rheumatoid arthritis(RA) based on network pharmacology.Methods: Through the traditional chinese medicine systems pharmacology database and analysis platform(TCMSP) and encyclopedia of traditional chinese medicine(ETCM),the main chemical components of Sishen Decoction were searched, screened and supplemented in combination with the literature.The smiles structural formula and 2 D structure of chemical components were retrieved through Pubchem organic small molecule bioactivity database.After screening by Swiss ADME database, the chemical components meeting the standard were uploaded toSwiss Target Prediction database to predict drug active targets.The targets related to RA diseases were obtained through human gene database(Gencards),online mendelian inheritance in man(OMIM),therapeutic target database(TTD) and DrugBank database.Wayne diagram was drawn and the common targets of active components of Sishen Decoction and RA were obtained.The protein-protein interaction network of drug and disease common targets was constructed based on String platform.Based on Metascape platform, gene ontology(go) biological function analysis and kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were carried out.Cytoscape 3.7 1 software was used to construct the "component-target-pathway" network, the network topology parameters of each component and target were obtained by using the function of network analyzer, and the important active components and core targets of Sishen Decoction in the treatment of RA were screened.Results: 60 active components and 353 related targets of Sishen Decoction were obtained by ETCM,TCMSP database and literature search;1 277 potential RA targets were obtained through GeneCards, OMIM,TTD and DrugBank;after mapping the action targets of drug components with RA related targets, 151 targets of Sishen Decoction for RA were obtained;2 322 items were obtained by GO biological function enrichment analysis, including 2089 biological processes, 79 cell components and 154 molecular functions;the enrichment analysis of KEGG pathway obtained 174 signal pathways, mainly involving interleukin-17(IL-17) signal pathway, tumor necrosis factor(TNF) signal pathway, Th17 cell differentiation, osteoclast differentiation and nuclear factor-kappa B(NF-κB) Signal path, etc;network topology analysis shows that the important active components of Sishen Decoction in the treatment of RA are quercetin, luteolin, kaempferol, wogonin and other compounds, and the important target genes are PTGS2,PTGS1,RELA,TNF,NFKB1,IL-6 and so on.Conclusion: Sishen Decoction may regulate the expression of target genes such as PTGS2,PTGS1,RELA,TNF,NFKB1 and IL-6 through active components such as quercetin, luteolin, kaempferol and wogonin, and then regulate biological pathways such as IL-17 signal pathway, TNF signal pathway, Th17 cell differentiation and osteoclast differentiation.