基于网络药理学与体外细胞实验探讨复方清痹片治疗类风湿性关节炎的作用机制

Mechanism of action of compound Qingbi tablets in treatment of rheumatoid arthritis:A study based on network pharmacology and in vitro experiment

目的:基于网络药理学与体外细胞实验探讨复方清痹片治疗类风湿性关节炎(RA)的作用机制。方法:基于TCMID、TCMSP和DrugBank数据库筛选复方清痹片的活性成分并预测潜在活性成分靶点;基于GeneCard、OMIM等数据库筛选RA疾病靶点;明确复方清痹片活性成分与RA的共同潜在靶标;利用STRING数据库和Cytoscape软件构建蛋白质-蛋白质相互作用(PPI)网络,并通过DAVID数据库进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析,RT-PCR检测复方清痹片对肿瘤坏死因子(TNF-α)诱导的人类风湿性关节炎成纤维样滑膜细胞(HFLS-RA)模型白介素6(IL-6)、TNF-α、丝氨酸/苏氨酸蛋白激酶1(AKT1)和血管内皮生长因子A(VEGFA)mRNA表达的影响。结果:筛选出217种潜在活性成分以及158个潜在靶点;IL-6、TNF、AKT1、白蛋白(ALB)、VEGFA等是复方清痹片治疗RA的关键靶点;复方清痹片主要通过炎性因子、趋化因子、免疫调控等信号通路来发挥治疗RA的作用。复方清痹片能够抑制HFLS-RA增殖,下调IL-6、TNF、AKT1和VEGFA mRNA的表达,这与网络药理学预测的结果相一致。结论:本研究揭示了复方清痹片治疗RA“活性成分-靶点-通路”的复杂关系,为后续该药的临床应用提供了研究基础。

Objective:To investigate the mechanism of action of compound Qingbi tablets in the treatment of rheumatoid arthritis(RA)based on network pharmacology and in vitro experiment.Methods:TCMID,TCMSP,and DrugBank databases were used to obtain the active components of compound Qingbi tablets and predict their potential targets,and GeneCard and OMIM databases were used to screen out the disease targets of RA;the common potential targets of the active components of compound Qingbi tablets and RA were clarified.STRING database and Cytoscape software were used to construct a protein-protein interaction network,and DAVID database was used to perform gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.RT-PCR was used to observe the effect of compound Qingbi tablets on the mRNA expression of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),serine/threonine protein kinase 1(AKT1),and vascular endothelial growth factor A(VEGFA)in a human rheumatoid arthritis fibroblast-like synoviocyte(HFLS-RA)model induced by TNF-α.Results:A total of 217 potential active components and 158 potential targets were screened out.IL-6,TNF,AKT1,albumin,and VEGFA were the key targets for compound Qingbi tablets in the treatment of RA,and compound Qingbi tablets exerted a therapeutic effect on RA mainly through the signaling pathways such as inflammatory factor,chemokine,and immunoregulation.Compound Qingbi tablets inhibited the proliferation of HFLS-RA and downregulated the mRNA expression of IL-6,TNF,AKT1,and VEGFA,which was consistent with the prediction results based on network pharmacology.Conclusion:This study reveals the complex“active component-target-pathway”relationship for compound Qingbi tablets in the treatment of RA,which provides a basis for the clinical application of this drug.