基于网络药理学及动物实验探讨桂苓四子方治疗痛风性肾病的作用机制

To investigate the mechanism of Guiling Sizi Decoction in the treatment of gouty nephropathy based on network pharmacology and animal experiment

目的利用网络药理学和动物实验研究桂苓四子方治疗痛风性肾病(GN)的疗效和作用机制。方法利用中药系统药理学数据库与分析平台(TCMSP)和SymMap数据库收集桂苓四子方的化学成分,结合TCMSP、SwissADME数据库及文献筛选活性成分,利用SwissTargetPrediction数据库预测成分作用靶点;通过OMIM、GeneCard数据库获取GN的靶点;利用STRING数据库建立成分-疾病共同靶点蛋白相互作用(PPI)网络,采用Cytoscape3.9.1软件对PPI网络进行筛选得出核心靶点;将活性成分-疾病共同靶点导入R Studio 4.2.3软件中,使用clusterProfiler软件包进行GO富集分析和KEGG通路富集分析。最后基于网络药理学结果,选择TNF/PI3K信号通路,通过体内实验进一步验证。结果共筛选出桂苓四子方492个活性成分,预测到潜在靶点1160个,其中278个与GN有关,PPI网络筛选得到关键靶点22个。GO和KEGG富集分析显示,桂苓四子方药可能通过干预TNF/PI3K通路改善GN。体内实验证实桂苓四子方有效降低了GN模型大鼠的SUA、Scr、BUN、尿蛋白水平及肾组织TNF-α、IL-1β、MCP-1水平,改善了肾组织炎症反应及纤维化,且显著下调了了大鼠肾组织中TNF-α、磷脂酰肌醇3′-激酶(PI3K)的表达。结论桂苓四子方药对可能通过干预TNF/PI3K通路改善GN的肾脏炎症反应,缓解肾纤维化。

Objective To investigate the efficacy and mechanism of Guiling Sizi Decoction in the treatment of gouty nephropathy(GN)by network pharmacology and animal experiment.Methods Chemical components of Guiling Sizi Decoction were collected using Traditional Chinese Medicine(TCM)Systematic Pharmacology Database and Analysis Platform(TCMSP)and SymMap Database.The active components of GN were screened by TCMSP,SwissADME database and literature.SwissTarget Prediction database was used to predict the targets of component action.The targets of GN were obtained by OMIM and GeneCard databases.The component-disease common target protein interaction(PPI)network was established using STRING database,and the core target was identified by Cytoscape3.9.1 software for screening PPI network.The active ingredientsdisease common targets were imported into R Studio 4.2.3 software,and the clusterProfiler software package was used for GO enrichment analysis and KEGG pathway enrichment analysis.Finally,based on the results of network pharmacology,TNF/PI3K signaling pathway was selected and further verified by in vivo experiments.Results A total of 492 active ingredients of Guiling Sizi Decoction were screened,and 1160 potential targets were predicted,of which 278 were related to GN,and 22 key targets were identified by PPI network screening.GO and KEGG enrichment analysis showed that Guiling Sizi Decoction may improve GN by interfering with TNF/PI3K pathway.In vivo experiments confirmed that Guiling Sizi Decoction effectively reduced the levels of SUA,Scr,BUN,urinary protein and TNF-α,IL-1βand MCP-1 in renal tissue of GN model rats,improved the inflammatory response and fibrosis of renal tissue,and significantly down-regulated the expression of TNF-αand phosphatidylinositol 3'-kinase(PI3K)in renal tissue of rats.Conclusion Guiling Sizi Decoction may improve the renal inflammatory response of GN and alleviate renal fibrosis by intervening with TNF/PI3K pathway.