网络药理学、分子对接结合实验验证探讨飞蛾藤治疗类风湿关节炎的作用机制  

Exploring mechanism of Porana racemosa Roxb.in treating rheumatoid arthritis based on integration of network pharmacology and molecular docking combined with experimental validation

作　　者：叶晨昱 李宁 陈音孜 曲彤 胡静 陈志永 任慧[2] YE Chen-yu;LI Ning;CHEN Yin-zi;QU Tong;HU Jing;CHEN Zhi-yong;REN Hui(School of Life Sciences,Northwest University,Xi'an 710069,China;Shaanxi Academy of Traditional Chinese Medicine,Xi'an 710003,China)

机构地区：[1]西北大学生命科学学院,陕西西安710069 [2]陕西省中医药研究院,陕西西安710003

出　　处：《药学学报》2025年第1期117-129,共13页Acta Pharmaceutica Sinica

基　　金：国家重点研发计划(2023YFC3504900);国家自然科学基金面上项目(81973419);陕西省中医药管理局“双链融合”中青年科研创新团队(2022-SLRH-YQ-003)。

摘　　要：通过网络药理学和分子对接技术,结合体外实验验证探究飞蛾藤(Porana racemosa Roxb.,PRA)治疗类风湿关节炎(rheumatoid arthritis,RA)的作用机制,为飞蛾藤治疗RA提供现代药理学依据。通过Swiss Target Prediction数据库预测已解析飞蛾藤中化学成分的潜在靶点;采用OMIM、GeneCards、TTD、Disgenet数据库检索RA的疾病靶点;利用STRING数据库和Cytoscape软件构建蛋白相互作用(PPI)网络和药材-成分-靶点网络;使用DAVID数据库进行GO(gene ontology)功能富集和KEGG(kyoto encyclopedia of genes and genomes)通路分析,并利用分子对接软件对飞蛾藤潜在活性成分与核心靶点进行对接;最后选取MH7A细胞进行细胞活力、划痕愈合、关键基因mRNA表达水平分析等实验,以探索飞蛾藤及其潜在活性成分对MH7A细胞增殖、迁移、凋亡的影响。该研究共筛选出飞蛾藤潜在活性成分靶点628个,RA靶点1890个,交集靶点235个,筛选出飞蛾藤治疗RA的主要潜在活性成分是咖啡酸乙酯、N-对反式香豆酰酪胺、亚麻酸甲酯等,核心靶点涉及肿瘤坏死因子(TNF)、基质金属蛋白酶9(MMP9)、前列腺素内过氧化物合酶2(PTGS2)等,富集分析确定了1200个GO功能富集条目和166条信号通路;分子对接结果显示,N-对反式香豆酰酪胺和咖啡酸乙酯与TNF、MMP9、半胱氨酸天冬氨酸蛋白酶3(CASP3)、PTGS2、B淋巴细胞瘤2(BCL2)均具有较好的结合活性;体外实验显示,飞蛾藤、咖啡酸乙酯和N-对反式香豆酰酪胺均可抑制MH7A细胞增殖、迁移和侵袭能力,上调凋亡相关基因CASP3 mRNA的表达,下调MMP9、PTGS2、BCL2 mRNA的表达,还能下调磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)mRNA和PI3K、p-AKT蛋白的表达。该研究初步揭示了飞蛾藤治疗RA可能与抑制MH7A细胞增殖、迁移和侵袭,诱导细胞凋亡,调控PI3K/AKT信号通路等有关。Through network pharmacology and molecular docking technology,combined with in vitro experiment verification,we explored the mechanism of action of Porana racemosa Roxb.(PRA)in the treatment of rheumatoid arthritis(RA),and provided a modern pharmacological basis for the treatment of RA by PRA.The potential target of chemical components in the analyzed moth rattan was predicted by Swiss Target Prediction database;OMIM,GeneCards,TTD and Disgenet databases were used to search the disease targets of RA;the protein interaction(PPI)network and medicine-composition-target network were constructed using STRING database and Cytoscape software;GO(gene ontology)functional enrichment and KEGG(kyoto encyclopedia of genes and genomes)pathway analysis were carried out using DAVID database,and molecular docking software was used to dock the potentially active ingredients of PRA and core targets;finally,MH7A cells were selected for cell viability,scratch healing and mRNA expression level analysis of key genes to explore the effects of PRA and their potentially active ingredients on the proliferation,migration and apoptosis of MH7A cells.In this study,a total of 628 potentially active ingredient targets,1890 RA targets and 235 intersection targets were identified.It was screened that the potentially active ingredients of RA treatment by PRA were ethylcaffeate,N-p-coumaroyltyramine,9,12,15-octadecatrienoic acid,methyl ester and so on,and the core targets involved tumor necrosis factor(TNF),matrix metalloproteinase 9(MMP9),prostaglandin-endoperoxide synthase 2(PTGS2)and so on.1200 GO entries and 166 KEGG pathway entries were obtained from the enrichment analysis;molecular docking results showed that N-p-coumaroyltyramine and ethylcaffeate had good binding activity with TNF,MMP9,cysteine-aspartate protease 3(CASP3),PTGS2,B-cell lymphoma 2(BCL2)proteins.In vitro experiments showed that PRA,ethylcaffeate and N-p-coumaroyltyramine could inhibit the proliferation,migration and invasion of MH7A cells,up-regulate the expression of apoptosi

关 键 词：类风湿关节炎 飞蛾藤 网络药理学 分子对接 凋亡 MH7A细胞 

分 类 号：R966[医药卫生—药理学]