金匮肾气丸防治骨质疏松症的机制研究

Study on the mechanism of Jinkuishenqi pills in the prevention and treatment of osteoporosis

目的探讨金匮肾气丸防治骨质疏松症的作用机制。方法从TCMSP数据库中筛选金匮肾气丸活性成分与作用靶标,用Cytoscape3.6构建金匮肾气丸“药物-活性成分-靶标基因”网络图;在DisGeNET、GeneCards数据库中查询骨质疏松症疾病基因,与金匮肾气丸活性成分靶标基因取交集后,从STRING数据库获取PPI网络信息,导入Cytoscape3.6构建网络图,再用CytoHubba插件中的MCC和Degree算法筛选并获得关键基因;用David对关键基因进行GO生物功能和KEGG通路富集分析,使用Omicshare绘制高级气泡图,通过KEGG数据库找出重要通路并标记关键基因。结果金匮肾气丸活性成分47种,对应196个作用靶标,与骨质疏松症疾病基因有145个交集靶标,筛选得到30个关键基因。GO富集分析结果表明,生物途径主要有药物反应、转录正调控和DNA模板化、RNA聚合酶Ⅱ启动子的转录正调控等;分子功能主要有酶、相同蛋白、蛋白等的结合;细胞组分为核浆、胞浆、细胞核等。关键基因则主要富集在TNF、MAPK、PI3K-Akt及其他重要信号通路上。结论金匮肾气丸中的槲皮素、豆甾淳、β-谷甾醇、山柰酚等核心活性成分通过TNF、MAPK、PI3K-Akt、HIF-1等信号通路干预AKT1、TP53、IL6、MAPK1、MAPK8、JUN等关键靶标基因的表达,促进成骨细胞生成、抑制破骨细胞分化,维持骨代谢平衡,从而达到防治骨质疏松症的目的。

Objective To investigate the mechanism of of Jinkuishenqi pills(JSP)in the prevention and treatment of osteoporosis.Methods The active components and targets of JSP were screened from TCMSP database.The network map of herb-ingredient-target of JSP was constructed using Cytoscape 3.6.The disease genes of osteoporosis were searched in DisGeNET and GeneCards online databases.After the intersection of these genes with the target genes of JSP,PPI network information was obtained from STRING database.Cytoscape 3.6 was used to construct the network diagram.David was used to perform the GO biological function and KEGG pathway enrichment analysis on key genes.Omicshare was used to draw the advanced bubble map.The key pathways were found and key genes were labeled using KEGG database.Results There were 47 active components corresponding to 196 target genes,145 intersecting target genes with osteoporosis disease genes,and 30 key genes were screened out from JSP.The result of GO enrichment analysis showed that the biological pathways mainly included response to drug,positive regulation of transcription,DNA templates,and positive regulation of transcription from RNA polymerase II promoter.The molecular functions mainly include enzyme binding,identical protein binding,and protein binding,etc.The cell components were divided into nucleoplasm,cytosol,and nucleus,etc.The key genes were mainly enriched in TNF,MAPK,PI3K-Akt,and other signaling pathways.Conclusion Quercetin,stigmasterol,beta-sitostero,kaempferol,and other core active ingredients in JSP intervene the expression of the key target genes AKT1,TP53,IL6,MAPK1,MAPK8,and JUN,through TNF,MAPK,PI3K-Akt,HIF-1 signaling pathways,promote osteoblast formation,inhibit osteoclast differentiation,and maintain the balance of bone metabolism,and achieve the purpose of prevention and treatment of osteoporosis.