基于网络药理学探讨新止骨增生丸治疗膝骨关节炎的作用机制

Mechanism of action of New Zhigu Zengsheng Pill in the treatment of osteoarthritis of the knee by using network pharmacology

目的 运用网络药理学方法探索新止骨增生丸治疗膝骨关节炎(KOA)的潜在作用机制。方法 利用中药系统药理学分析平台(TCMSP)数据库筛选新止骨增生丸的活性成分及其作用靶点;利用GeneCards、OMIM和DisGeNet数据库检索KOA疾病的相关靶点;通过Venny 2.1.0在线平台对新止骨增生丸的活性成分作用靶点与KOA疾病相关靶点映射取交集,得到新止骨增生丸治疗KOA的关键靶点(共同靶点);将共同靶点导入String平台构建蛋白相互作用(PPI)网络,并将结果导入Cytoscape 3.7.1软件进行拓扑分析,提取核心靶点;运用注释、可视化和集成发现(DAVID)数据库对核心靶点进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,并利用Cytoscape 3.7.1软件构建“活性成分-关键靶点-通路”网络。结果 共获得活性成分54个,对应靶点262个,疾病靶点2459个,共同靶点142个,经过蛋白互作及拓扑分析,得到核心靶点13个, GO功能富集分析得到821个生物过程(BP), 94个细胞组分(CC), 139个分子功能(MF)和167条通路(P<0.05)。将“活性成分-核心靶点-通路”网络图进行拓扑分析,共筛选出槲皮素、β-谷甾醇、木犀草素、山柰酚等12个核心成分,以及肿瘤坏死因子(TNF)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、白细胞介素-6(IL-6)、血管内皮生长因子A (VEGFA)、白细胞介素-1β(IL-1β)等16个关键靶点。结论 新止骨增生丸可能通过槲皮素、β-谷甾醇、山柰酚等主要活性成分,作用于AKT1、TNF、VEGFA等核心靶点,通过高级糖基化终末产物-受体(AGE-RAGE)、TNF等信号通路,协同发挥对KOA的治疗作用。

Objective To explore the potential mechanism of action of New Zhigu Zengsheng Pill in the treatment of osteoarthritis of the knee(KOA)by using network pharmacology.Methods The active constituents and their targets of New Zhigu Zengsheng Pill were screened by traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)database.GeneCards,OMIM and DisGeNet databases were used to search KOA disease-related targets.Through Venny 2.1.0 online platform,the intersection mapping between the active ingredient target of New Zhigu Zengsheng Pill and the disease-related target of KOA was obtained to obtain the key target(common target)of New Zhigu Zengsheng Pill for treating KOA.The common targets were imported into the String platform construction protein-protein interaction(PPI)network,and the results were imported into Cytoscape 3.7.1 software for topological analysis to extract the core targets.Gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Gnomes(KEGG)pathway enrichment analysis were performed on the core targets by using Database for Annotation,Visualization and Integrated Discovery(DAVID)database.The"Active ingredient-key target-pathway"network was constructed by using Cytoscape 3.7.1 software.Results A total of 54 active ingredients were obtained,corresponding to 262 targets,2459 disease targets and a total of 142 targets.Through protein interaction and topological analysis,13 core targets were obtained,and 821 biological processes(BP),94 cell components(CC),139 molecular functions(MF)and 167 pathways were obtained by GO functional enrichment analysis(P<0.05).Topological analysis of the network diagram of"active ingredient-core target-pathway"was performed,and 12 core components such as quercetin,β-sitosterol,luteolin,kaempferol,and 16 key targets such as tumor necrosis factor(TNF),AKT serine/threonine kinase 1(AKT1),interleukin-6(IL-6),vascular endothelial growth factor A(VEGFA),interleukin-1β(IL-1β),and so on were selected.Conclusion New Zhigu Zengsheng Pill may act on AKT1,TNF,VEGFA and other core targets through quercetin,β-sitosterol,kaempferol and other active components,and play a synergistic role in the treatment of KOA through advanced glycation end products-receptor for advanced glycation end products(AGE-RAGE),TNF and other signaling pathways.