西洋参-红花有效成分治疗早期糖尿病肾病的网络药理学和实验验证

Network Pharmacology and Experimental Validation of Effective Components of Panacis Quinquefolii Radix and Carthamus tinctorius in Treatment of Early Diabetic Nephropathy

目的:通过网络药理学、分子对接技术及细胞实验探讨西洋参-红花有效成分治疗早期糖尿病肾病(DN)的作用机制。方法:运用中药系统药理学数据库与分析平台(TCMSP)获得西洋参-红花活性成分的干扰靶点,再与GeneCards、TTD等数据库中早期DN基因靶点相映射即为交集靶点。采用STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络,运用Cytoscape 3.8.2软件筛选出核心靶点;通过DAVID数据库进行基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)富集分析,运用Cytoscape 3.8.2软件进行“有效成分-关键靶点-通路-疾病”网络图展示;通过Autodock Tools软件进行分子对接,采用MTT法检测有效成分对脂多糖(LPS)诱导的人肾小球系膜细胞(HMC)活性,利用酶联免疫吸附试验检测有效成分对炎症介质的影响。结果:得到西洋参-红花治疗早期DN的活性成分有24个,对应的药物靶点207个,其中72个和早期DN相关。网络拓扑参数分析得出了肿瘤坏死因子(TNF)、白细胞介素-6(IL-6)、IL-1β等15个关键靶点,KEGG通路分析筛选出晚期糖基化终末产物(AGE)-AGE受体(RAGE)、TNF、低氧诱导因子-1(HIF-1)、IL-17、核因子κB等20个信号通路;分子对接表明IL-6与槲皮素、木犀草素、黄芩素、人参皂苷rh2稳定结合。细胞实验得出西洋参-红花活性成分槲皮素、木犀草素、黄芩素能显著抑制IL-6、TNF-α和IL-1β表达。结论:本研究初步证实西洋参-红花有效成分能有效抑制早期DN炎症,对肾脏具有保护作用,为后期深入研究提供了依据。

Objective:To investigate the mechanism of effective components of Panacis Quinquefolii Radix and Carthamus tinctorius in the treatment of early diabetic nephropathy(DN) through network pharmacology,molecular docking,and cell experiments.Methods:The interference targets of the active components of Panacis Quinquefolii Radix and Carthamus tinctorius were obtained by using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and then mapped with the early DN gene targets in GeneCards,TTD,and other databases to form intersection targets.The protein-protein interaction(PPI) network was constructed by the STRING database,and the core targets were selected by Cytoscape 3.8.2 software.Gene ontology(GO) enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analysis were performed by the DAVID database,and then the network diagram of “effective component-key target-pathway-disease” was presented by Cytoscape 3.8.2 software.Molecular docking was carried out using Autodock Tools.MTT assay was used to detect the activity of effective components on LPS-induced human mesangial cells(HMCs),and ELISA was used to detect the effect of effective components on inflammatory factors.Results:There were 24 active components and 207 corresponding drug targets of early DN treated with Panacis Quinquefolii Radix and Carthamus tinctorius,of which 72 were associated with early DN.Network topological parameter analysis yielded 15 key targets such as TNF,IL-6,and IL-1β,and KEGG pathway analysis selected 20 signaling pathways such as AGE-RAGE,TNF,HIF-1,IL-17,and NF-κB;molecular docking indicated that IL-6 stably bound to quercetin,luteolin,baicalein,and ginsenoside rh2.Cell experiments found that quercetin,luteolin,and baicalein could significantly inhibit the expression of IL-6,TNF-α,and IL-1β.Conclusion:The effective components of Panacis Quinquefolii Radix and Carthamus tinctorius can effectively inhibit the inflammation of early DN and protect the kidney.These results provide a basis for subsequent in-depth study.