黄芪及其活性成分的利水作用机制探讨

Discussion on Water Diuresis Mechanism of Huangqi (Radix Astragali seu Hedysari) and Active Components

目的:探讨黄芪利尿的活性成分及作用机制。方法:基于中药系统药理学数据库与分析平台、PubChem数据库及Swiss Target Prediction数据库检索黄芪的化学成分及相关作用靶点。在人类基因数据库与人类孟德尔遗传数据库预测水肿疾病靶点,与黄芪活性成分相关作用靶点进行映射,并绘制韦恩图,获取黄芪治疗水肿的潜在靶点。将黄芪活性成分与潜在靶点导入Cytoscape 3.7.2软件,构建“活性成分-潜在靶点”相互作用网络。将潜在靶点导入STRING数据库构建PPI网络并筛选核心基因。将黄芪活性成分相关作用靶点导入DAVID数据库进行基因本体(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。选取54只6~8周龄SPF级SD雄性大鼠进行水负荷实验,分别收集药物干预后0~2 h、2~4 h、4~6 h、7~24 h内的大鼠尿液,考察黄芪及其活性成分的利尿作用。结果:共得到25个黄芪活性成分及248个相关靶点,710个水肿相关靶点,黄芪治疗水肿的潜在靶点79个;通过“活性成分-水肿靶点”相互作用网络获得芒柄花素、毛蕊异黄酮等15个活性成分;通过PPI网络获得41个核心靶点;GO功能富集分析共获得生物过程464个条目,分子功能77个条目及细胞组分45个条目;KEGG通路富集分析共得到106条通路。大鼠水负荷实验显示,与正常组比较,灌胃后0~2 h、2~4 h及0~6 h内呋塞米组、黄芪小剂量组及毛蕊异黄酮组的尿量显著增多;灌胃后4~6 h内呋塞米组、黄芪小剂量组、黄芪大剂量组及芒柄花素组的尿量显著增多;灌胃后7~24 h内黄芪小剂量组、毛蕊异黄酮组大鼠的尿量明显增加,差异均具有统计学意义。结论:黄芪可通过多成分、多靶点、多通路发挥利尿作用,且小剂量黄芪利尿作用更为显著。

Objective:To explore the diuretic active components of Huangqi and its mechanism of action.Methods:Based on the traditional Chinese medicine system pharmacology database and analysis platform,PubChem database,and Swiss Target Prediction database,the chemical constituents and related active targets of Huangqi were searched.The edema disease targets were predicted in the human gene database and the human Mendelian genetic database,the targets related to the active components of Huangqi were mapped,and Venn diagrams were drawn to obtain the potential targets of Huangqi in the treatment of edema.Then the active components and potential targets of Astragalus were imported into Cytoscape 3.7.2 software to construct an "active component-potential target" interaction network.Import potential targets into the STRING database to construct a PPI network and screen core genes.The relevant targets of the active components of Huangqi were imported into the DAVID database for gene ontology(GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis.And 54 6-8 week-old SPF SD male rats were selected for the water load experiment,and the rat urine within 0-2 h,2-4 h,4-6 h,and 7-24 h after drug intervention was collected,to investigate the diuretic effect of Huangqi and its active components.Results:A total of 25 active components of Huangqi and 248 related targets,710 edema-related targets,and 79 edema-treating potential targets of Huangqi were obtained.15 active ingredients such as formononetin and mullein through the "active component-edema target" interaction network were obtained.41 core targets were obtained through the PPI network.A total of 464 items of biological process,77 items of molecular function,and 45 items of cellular components were obtained through GO functional enrichment analysis.And a total of 106 pathways were obtained from the KEGG pathway enrichment analysis.The water load test of rats showed that compared with the normal group,the urine output in the furosemide group,the low-dose Huangqi group,and the mullein isoflavone group increased significantly at 0-2 h,2-4 h,and 0-6 h after gavage.The urine output in the furosemide group,low-dose Huangqi group,high-dose Huangqi group,and formononetin group increased significantly within 4-6 h after gavage.And within 7-24 h after gavage,the urine output of the rats in the low-dose Huangqi group and the mullein group was significantly increased,and the differences were statistically significant.Conclusion:Astragalus can exert a diuretic effect through multiple components,multiple targets,and multiple pathways,and the diuretic effect of low-dose Huangqi is more significant.