基于网络药理学探究加味独活寄生合剂对膝骨关节炎作用机制及实验验证

Mechanism and experimental verification of jiawei duhuo jisheng mixture in treatment of knee osteoarthritis based on network pharmacology

目的使用网络药理学探究加味独活寄生合剂对膝骨关节炎(knee osteoarthritis,KOA)的可能作用机制,并通过动物实验验证。方法检索TCMSP、BATMAN-TCM数据库获取加味独活寄生合剂中各药物有效化学成分及作用靶点,在GEO、GeneCard、OMIM 3个数据库中检索获得KOA疾病靶点,将药物作用靶点与疾病靶点取交集获得共同靶点,随后通过STRING数据库及Cytoscape3.7.2软件对共同靶点进行构建“成分-疾病靶点网络图”及蛋白互作网络分析,借助Metascape数据库行GO、KEGG富集分析,最后动物实验对关键靶点予以验证。结果共筛选出加味独活寄生合剂治疗KOA有效活性成分180种,作用于152个共同靶点。PPI网络分析显示AKT1可能是核心靶点;GO、KEGG富集分析主要涉及炎症反应、凋亡信号通路等。动物实验证实,加味独活寄生合剂后能够改善KOA兔模型的骨关节炎严重程度,下调血清及关节液中IL-1β、TNF-α、Caspase 3、BAX表达水平,下调关节软骨中AKT1mRNA及蛋白表达水平。结论加味独活寄生合剂可能通过多种成分、多个靶点,涉及多条分子通路发挥抗炎、抗凋亡的作用,从而达到治疗KOA的作用。

Aim To explore and verify the possible mechanism of Jiawei Duhuo Jisheng Mixture(JDJM)in the treatment on Knee Osteoarthritis(KOA)via using network pharmacology and animal experiment.Methods The ingredients of JDJM and relevant targets were collected from TCMSP and BATMAN-TCM database.The KOA-related targets were collected from GeneCard,OMIM and GEO databases.The common targets were acquired by intersecting ingredients-related and KOA-related targets,and then the Ingredient-Disease-Target Network and PPI network were constructed by Cytoscape 3.7.2 software and STRING platform.GO and KEGG enrichment analysis were performed based on Metascape database.Finally,the key targets and relevant mechanism were validated via animal experiment.Results In the network pharmacology study,180 active ingredients related to treatment on KOA by JDJM were collected,and 152 common targets were confirmed.PPI network analysis showed that AKT1 might be the key targets of JDJM in the treatment on KOA.GO and KEGG enrichment analysis revealed that the key target mainly concentrated on inflammatory response and apoptosis.Animal experiment confirmed that JDJM could improve lesion in KOA rabbits,and suppress the expression levels of IL-1β,TNF-α,Caspase 3 and BAX in serum and articular fluid.AKT1 expression(including mRNA and protein)in articular cartilage was also down-regulated.Conclusions Based on the results of network pharmacology and animal experiment,JDJM may relieve KOA severity by anti-inflammatory and anti-apoptotic effects through a variety of molecular signaling pathways.