基于网络药理学探讨托法替布联合甲氨蝶呤治疗类风湿关节炎的作用机制

Systematic elaboration of the mechanism of tofacitinib combined with methotrexate in the treatment of rheumatoid arthritis via network pharmacology

目的:探讨托法替布联合甲氨蝶呤治疗类风湿关节炎(RA)的分子靶点和作用机制。方法:通过PharmMapper平台获取托法替布和甲氨蝶呤的预测药效团及其主要元素,基于STRING平台分析靶蛋白之间的相互作用关系(PPI),采用Cytoscape 3.7.2软件进行药物的预测靶点可视化分析,并通过GO富集分析和KEGG通路分析探究其治疗RA的可能分子机制。结果:通过PharmMapper平台共获得137个托法替布和甲氨蝶呤的预测靶蛋白/基因,PPI结果证实Janus激酶(JAK)2、JAK3、信号转导与转录激活因子(STAT)1、蛋白激酶B(AKT)1、白细胞介素(IL)-2、丝裂原活化蛋白激酶(MAPK)1、MAPK8、表皮生长因子受体(EGFR)等为主要预测靶蛋白/基因,GO分析表明上述靶蛋白/基因在受损DNA结合(P=0.002)、ATP酶结合(P=0.002)、核受体活性(P=0.002)、转录因子活性(P=0.002)、直接配体调控序列特异性DNA结合(P=0.002)、类固醇激素受体活性(P=0.004)、生长因子受体结合(P=0.022)和内肽酶活性(P=0.029)等功能处显著富集,结合KEGG通路分析证实托法替布联合甲氨蝶呤与JAK-STAT、转化生长因子(TGF)-β、磷脂酰肌醇3激酶(PI3K)/-AKT、p53、Ras、MAPK、血管内皮生长因子(VEGF)、核因子(NF)-κB、Notch等信号通路密切相关。结论:托法替布联合甲氨蝶呤治疗RA的分子机制具有多基因、多靶点的特点,并通过抑制炎症因子、血管生成、细胞自噬等方式调节免疫性疾病。

Objective:To explore the molecular target proteins/genes and mechanism of tofacitinib combined with methotrexate in the treatment of rheumatoid arthritis(RA).Methods:The predicted pharmacophore and characteristics of tofacitinib and methotrexate were obtained through PharmMapper platform.The protein-protein interaction(PPI)network of target proteins/genes was analyzed based on STRING platform.The network visualization of target proteins/genes of drugs was performed by Cytoscape 3.7.2 software,and the molecular mechanisms of two drugs to treat RA were further explored through GO enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Results:A total of 137 tofacitinib and methotrexate target proteins/genes were obtained via PharmMapper platform.PPI results confirmed that Janus kinase(JAK)2,JAK3,signal transduction and transcriptional activators(STAT)1,protein kinase B(AKT)1,interleukin(IL-2),mitogen activated protein kinase(MAPK)1,MAPK8 and epidermal growth factor receptor(EGFR)were the main predicted target proteins/genes.GO analysis showed that these target proteins/genes were significantly enriched in impaired DNA binding(P=0.002),ATPase binding(P=0.002),nuclear receptor activity(P=0.002),transcription factor activity(P=0.002),direct ligand regulatory sequence specific DNA binding(P=0.002),steroid hormone receptor activity(P=0.004),growth factor receptor binding(P=0.022)and endopeptidase activity(P=0.029).Combined with KEGG pathway analysis,tofatib combined with methotrexate,JAK-STAT signaling pathway,transforming growth factor(TGF)-βsignaling pathway,phosphatidylinositol 3 kinase(PI3K)-AKT signaling pathway,p53 signaling pathway,Ras signaling pathway,MAPK signaling pathway,vascular endothelial growth factor(VEGF)signaling pathway,nuclear factor(NF)-κB signaling pathway,Notch signaling pathway and other signaling pathways were closely related.Conclusion:The molecular mechanism of tofacitinib combined with methotrexate in the treatment of RA has the multi-gene and multi-target characteristics,and regulates immune diseases by inhibiting inflammatory factors,angiogenesis,autophagy and other functions.