基于网络药理学和体内实验研究竹叶石膏汤治疗痛风性关节炎的分子机制

Investigation on the Molecular Mechanism of Zhuye Shigao Decoction(竹叶石膏汤)in the Treatment of Gouty Arthritis Based on Network Pharmacology and In Vivo Experiments

目的:通过生物信息学技术及分子对接技术挖掘竹叶石膏汤治疗痛风性关节炎(GA)的分子机制,并通过动物实验进行验证。方法:利用公共数据库TCMSP、Batman-TCM、DrugBank和UniProt搜索竹叶石膏汤的有效活性成分及相应靶蛋白,并基于GeneCards、OMIM数据库查询GA相关靶点。分别利用R语言中Venn包,以及Cytoscape中的BisoGenet插件预测竹叶石膏汤治疗GA的潜在分子,利用Cytoscape软件构建竹叶石膏汤-活性成分-靶点交互网络及竹叶石膏汤-痛风性关节炎-靶蛋白核心网络。通过网络拓扑学分析筛选竹叶石膏汤治疗GA的核心靶分子,并对核心基因进行GO、KEGG富集分析。通过动物实验验证竹叶石膏汤治疗GA的核心靶点。结果:共筛选出147种竹叶石膏汤治疗GA的潜在有效活性化合物,挖掘了223个与GA相关的潜在靶点,计算得出40个核心靶蛋白。GO功能富集分析显示,核心靶点基因的BP功能主要包括炎症反应的调节、细胞对脂多糖的反应、细胞对细菌来源分子的反应等,CC的靶点主要富集在细胞质囊泡腔、囊泡腔、质膜筏等,MF功能主要涉及细胞因子活性、细胞因子受体结合、受体配体活性等。这些核心靶基因参与IL-17信号通路、TNF信号通路、糖尿病并发症中的AGE-RAGE信号通路等与痛风性关节炎发生、发展相关的信号通路。动物实验发现竹叶石膏汤可有效抑制GA大鼠滑膜组织IRAK4 mRNA及IL-6 mRNA的表达,减缓踝关节肿胀,降低血清尿酸水平。结论:竹叶石膏汤可通过“多成分-多靶点-多途径”有效抑制滑膜组织IRAK4 mRNA、IL-6 mRNA的表达,减缓踝关节肿胀,降低血清尿酸,从而发挥对GA的治疗作用。

Objective:To uncover the molecular mechanism of Zhuye Shigao Decoction for the treatment of gouty arthritis(GA)by bioinformatics and molecular docking technology,and to validate it by animal experiments.Methods:TCMSP,Batman-TCM,DrugBank and UniProt were used to search the effective active ingredients and corresponding target proteins of Zhuye Shigao Decoction,and the GA-related targets were searched based on GeneCards and OMIM databases.The Venn package in R language and the BisoGenet plug-in in Cytoscape were used to predict the potential molecules of Zhuye Shigao Decoction for GA treatment,respectively,and the Cytoscape software was used to construct the Zhuye Shigao Decoction-active compounds-target interaction network and the Zhuye Shigao Decoction-GA-target protein core network.The core target molecules of Zhuye Shigao Decoction for GA treatment were screened by network topology analysis,and the core genes were analyzed by GO and KEGG enrichment.Finally,the core targets of Zhuye Shigao Decoction for GA treatment were verified by animal experiments.Results:A total of 147 potentially effective active compounds of Zhuye Shigao Decoction for GA treatment were screened,223 potential targets related to GA were mined,and 40 core target proteins were calculated.GO functional enrichment analysis showed that the core target genes mainly enriched in regulation of inflammatory response,cellular response to lipopolysaccharide,cellular response to molecules of bacterial origin,etc in BP,cytoplasmic vesicle lumen,vesicle lumen,and plasma membrane rafts,etc in CC,and cytokine activity,cytokine receptor binding,and receptor ligand activity in MF.These core target genes enriched in IL-17 signaling pathway,TNF signaling pathway,AGE-RAGE signaling pathway in diabetic complications and other signaling pathways related to the occurrence and development of GA.Animal experiments revealed that Zhuye Shigao Decoction could effectively inhibit the expression of IRAK4 mRNA and IL-6 mRNA in synovial tissue of GA rats,slow down ankle joint swelling and reduce serum uric acid level.Conclusion:Zhuye Shigao Decoction can effectively inhibit the expression of IRAK4 mRNA and IL-6 mRNA in synovial tissues,reduce ankle swelling and lower serum uric acid through"multi-component-multi-target-multi-pathway",thus exerting the therapeutic effect on GA.