应用网络药理学探究壮医清毒伸筋方治疗类风湿关节炎的作用机制

Mechanism of Qingdu Shenjin Formula in the Treatment of Rheumatoid Arthritis Based on Network Pharmacology

目的:网络药理学探究壮医清毒伸筋方治疗类风湿关节炎(RA)的作用机制。方法:检索中药系统药理学数据库与分析平台(TCMSP)、人类在线孟德尔遗传数据库(OMIM)、人类基因数据库(GeneCards)获取清毒伸筋方与RA交集靶点。网络可视化和分析软件(Cytoscape)获取主要活性成分,归一化相互匹配法(NCC)计算核心靶点。对核心靶点进行基因本体(GO)富集分析、京都基因与基因组百科全书(KEGG)富集分析,用分子对接分析主要活性成分与核心靶点的结合力。构建胶原诱导性关节炎(CIA)大鼠模型验证主要蛋白浓度。结果:获得145种活性成分和238个药靶点,交集靶点151个。富集分析提示交集靶点涉及19个GO富集分析条目和晚期糖基化终末产物(AGEs)-晚期糖基化终末产物受体(RAGE)信号通路、肿瘤坏死因子(TNF)信号通路等75条信号通路。主要活性成分为槲皮素、柄花黄素、豆甾醇、木犀草素、β-谷甾醇;分子对接结果显示它们与前列腺素内过氧化物合酶2(PTGS2)、基质金属蛋白酶9(MMP9)、丝氨酸/苏氨酸蛋白激酶(AKT1)等蛋白具有良好的结合性。动物实验验证,清毒伸筋方有效减轻CIA大鼠关节肿胀程度,显著降低血清MMP9、AKT1和PTGS2浓度。结论:清毒伸筋方发挥多成分、多靶点、多通路特点作用于RA的炎症级联反应。

Objective:To explore the mechanism of Zhuang ethnomedicine Qingdu Shenjin Formula in the treatment of rheumatoid arthritis(RA) through network pharmacology.Methods:The intersection targets of Qingdu Shenjin Formula and RA were identified using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),the Online Mendelian Inheritance in Man(OMIM) database,and the GeneCards database.Major active components were obtained using network visualization and analysis software(Cytoscape),and core targets were calculated using the normalized cross-correlation(NCC) method.Gene Ontology(GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were conducted on the core targets.Molecular docking was used to analyze the binding affinity between major active components and core targets.The collagen-induced arthritis(CIA) rat model was constructed to verify the concentration of major proteins.Results:A total of 145 active components and 238 drug targets were identified,with 151 intersection targets.Enrichment analysis indicated that the intersection targets were involved in 19 GO terms and 75 signaling pathways,including the advanced glycation end products(AGEs)-receptor for AGEs(RAGE) signaling pathway and the tumor necrosis factor(TNF) signaling pathway.The main active components were quercetin,anthocyanin,stigmasterol,luteolin,and β-sitosterol.Molecular docking results showed good binding affinity between these components and proteins such as prostaglandin-endoperoxide synthase 2(PTGS2),matrix metalloproteinase 9(MMP9),and serine/threonine kinase(AKT1).Animal experiments confirmed that Qingdu Shengjin Formula significantly reduced joint swelling in CIA rats and lowered serum concentrations of MMP9,AKT1,and PTGS2.Conclusion:Qingdu Shengjin Formula exerts its effects on the inflammatory cascade reaction of RA through multiple components,multiple targets,and multiple pathways.