基于网络药理学和实验研究探讨尿感合剂治疗阴道炎的作用机制

Mechanism of Niaogan Mixture in the Treatment of Vaginitis Based on Network Pharmacology and Experimental Validation

目的探讨经典名方“尿感合剂”治疗阴道炎的作用机制。方法在TCMSP数据库中获取“尿感合剂”中主要的7味药材(泽泻、茯苓、金钱草、凤尾草、萹蓄、海金沙、甘草)的活性成分和对应的靶基因。由Uniprot网站根据药物靶基因获取标准靶蛋白名称。借助GeneCards(The Human Gene Database)数据库和人类孟德尔遗传综合数据库(Online Mendelian Inheritance in Man,OMIM)筛选阴道炎疾病靶基因。基于VENNY分析原理预测“尿感合剂”药物治疗阴道炎的潜在治疗靶点基因。由String数据库获取潜在治疗靶点的蛋白质互作关系,蛋白质-蛋白质相互作用关系由Cytoscape软件进行可视化处理构建PPI网络图。利用DAVID数据库对潜在治疗靶点进行GO富集分析和KEGG通路富集分析,借助生信在线工具作图。借助分子对接技术进行验证。通过体外抑菌实验研究不同浓度的“尿感合剂”对阴道炎疾病中常见的三种标准菌株(大肠埃希菌、金黄色葡萄球菌、粪肠球菌)作用疗效。结果通过VENNY分析获得潜在治疗靶点118个,筛选活性成分74个,预测有效靶点包括TP53、RELA、MAPK1、AKT1、IL6等。GO富集分析显示涉及27个生物过程,涉及细胞组分14个,涉及分子功能8个,KEGG通路富集获得230条通路。体外实验结果显示随着药物浓度的递减抑菌作用也随着降低,表明“尿感合剂”具有抑菌作用且具有一定的药物浓度依赖性。结论经典名方“尿感合剂”以多成分、多靶点和多通路的协同作用发挥治疗阴道炎的疗效。

Objective To explore the mechanism of classic prescription Niaogan Mixture in the treatment of vaginitis.Methods The active components and corresponding target genes of seven drugs[Zexie(Alismatis Rhizoma),Fuling(Poria),Jinqiancao(Lysimachiae Herba),Fengweicao(Pteridis Multifidae Herba),Bianxv(Polygoni Avicularis Herba),Haijinsha(Lygodii Spora),and Gancao(Glycyrrhizae Radix et Rhizoma)]in Niaogan Mixture were obtained from TCMSP.Standard target protein names were obtained from Uniprot website based on drug target genes.The target genes of vaginitis were screened out from The Human Gene Database(GeneCards)and Online Mendelian Inheritance in Man(OMIM).The potential therapeutic target genes of Niaogan Mixture in the treatment of vaginitis were predicted based on VENNY analysis.Protein-protein interaction(PPI)relationship for potential therapeutic targets was obtained from the String database and visualized by Cytoscape software to construct a PPI network.GO enrichment and KEGG pathway enrichment analyses of potential therapeutic targets were performed using the DAVID database,and BioLadder online tool was used for plotting.Validation was performed with the aid of molecular docking technique.In vitro antibacterial test was used to analyze the effect of different concentrations of Niaogan Mixture on three standard strains(Escherichia coli,Staphylococcus aureus,and Enterococcus faecalis)commonly found in vaginitis.Results A total of 118 potential therapeutic targets were obtained by VENNY analysis and 74 active components were screened out.The predicted effective targets included TP53,RELA,MAPK1,AKT1,and IL6.GO enrichment analysis showed 27 biological processes,14 cellular components,and eight molecular functions.KEGG pathway enrichment yielded 230 pathways.The results of the in vitro experiment showed that the antibacterial effect also decreased with the decrease in drug concentration,indicating that Niaogan Mixture had an antibacterial effect in a concentration-dependent manner.Conclusion The classic prescription Niaogan Mixture plays an effective role in the treatment of vaginitis with the synergistic effect of multiple components,multiple targets,and multiple pathways.