基于网络药理学和分子对接的黄蜀葵花干预糖尿病肾病的作用机制研究

Based on the network pharmacology and molecular docking method to explore the mechanism of flowers of Abelmoschus manihot in the intervention of diabetic kidney disease

目的运用网络药理学和分子对接的方法研究黄蜀葵花干预糖尿病肾病的多成分、多靶点、多通路作用机制,旨在为其基础研究及临床应用提供依据。方法通过ETCM、HERB数据库结合文献报道,筛选黄蜀葵花活性成分,并通过PharmMapper数据库预测其潜在靶点,建立靶点数据库。通过Genecards、OMIM、Drugbank、TTD等数据库筛选糖尿病肾病相关靶点,应用R4.1.0软件取两者交集靶点。通过STRING平台获得蛋白互作(PPI)关系,并应用Cytoscape 3.8.2构建PPI网络并分析其中关键靶点。通过David数据库进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)基因富集分析,得到黄蜀葵花干预糖尿病肾病的潜在作用通路,采用Cytoscape 3.8.2构建"成分-作用靶点-通路"调控网络,使用AutoDock Vina 1.1.2软件进行分子对接验证。结果从黄蜀葵花中筛选出43个活性成分,其中涉及糖尿病肾病作用靶点44个,Cytoscape分析最终筛选出9个关键靶点。GO富集分析结果显示,根据P值<0.05,筛选出生物过程(BP)120个,细胞组分(CC)23个,分子功能(MF)37个。KEGG富集分析结果显示,黄蜀葵花主要作用于肿瘤坏死因子(TNF)、丝裂原活化蛋白激酶(MAPK)等信号通路。分子对接结果显示,大部分靶点与成分的结合活性较好。结论基于网络药理学探讨了黄蜀葵花干预糖尿病肾病多成分-多靶点-多通路的作用特点,预测黄蜀葵花主要活性成分槲皮素-3-O-刺槐糖苷、芦丁、槲皮素-3’-O-葡萄糖苷、银椴苷等,可能通过白蛋白(ALB)、表皮生长因子受体(EGFR)、雌激素受体(ESR1)、过氧化物酶体增值因子活化受体(PPARG)、凝血酶原(prothrombin,F2)等靶点,作用于肿瘤坏死因子TNF、MAPK等信号通路干预糖尿病肾脏病,为进一步开展黄蜀葵花干预糖尿病肾病作用机制研究提供了新的思路和方法。

Objective Using the methods of network pharmacology and molecular docking, the mechanism of multicomponent,multitarget and multipathway action of flowers of Abelmoschus manihot in the intervention of diabetic kidney disease was studied,aiming to provide basis for its basic research and clinical application. Methods ETCM and HERB database combined with literature reports were used to screen the active ingredients of flowers of Abelmoschus manihot, and PharmMapper database was used to predict its potential targets and establish target database. Genecards, OMIM, Drugbank, TTD and other databases were used to screen the related targets of diabetic kidney disease, and R4.1.0 software was used to select the intersection targets. Protein interaction(PPI)relationships were obtained using STRING platform, and PPI networks were constructed using Cytoscape3.8.2 and the key targets were analyzed. David database was used for gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) gene enrichment analysis to obtain the potential pathway of flowers of Abelmoschus manihot in the intervention of diabetic kidney disease. Cytoscape 3.8.2 was used to construct the "components-target-pathway" regulatory network. AutoDock Vina 1.1.2 software was used for molecular docking verification. Results Forty-three active components were screened from flowers of Abelmoschus manihot, including 44 targets of diabetic kidney disease, and 9 key targets were screened by Cytoscape analysis. According to the results of GO enrichment analysis, 120 biological processes, 23 cell components, and 37 molecular functions were screened out according to P < 0.05. KEGG enrichment analysis showed that flowers of Abelmoschus manihot mainly acted on tumor necrosis factor(TNF) and mitogen-activated protein kinase(MAPK) signaling pathways. The results of molecular docking showed that most of the targets had good binding activity with the compounds. Conclusion Based on network pharmacology, the effects of multicomponent, multitarget and multipathway on the flowers of Abelmoschus manihot in the intervention of diabetic kidney disease were discussed. The main active components, such as quercetin-3-O-robinobioside, rutin, quercetin 3’-O-glucoside, and tiliroside may act on TNF, MAPK and other signaling pathways through ALB, EGFR, ESR1, PPARG and F2 to intervene diabetic nephropathy. This study provided a new idea and method for further study on the mechanism of the intervention on diabetic kidney disease.