基于网络药理学及细胞实验探讨丹蛭降糖胶囊干预糖尿病肾脏病的作用机制

Exploring the mechanism of action of Danzhi Jiangtang Capsule in diabetic nephropathy based on network pharmacology and cellular experiments

关键词:  丹蛭降糖胶囊  糖尿病肾脏病  网络药理学  肾小管上皮细胞  PI3K/Akt信号通路

Key words:  Danzhi Jiangtang Capsule  diabetic kidney disease  Network Pharmacology  renal tubular epithelial cell  PI3K/AKT signaling pathway

摘要:

[目的] 利用网络药理学结合细胞实验探讨丹蛭降糖胶囊干预糖尿病肾脏病(DKD)的作用机制。[方法] 通过中药系统药理学数据库分析平台(TCMSP)及本草组鉴(HERB)数据库收集丹蛭降糖胶囊有效成分,利用PubChem数据库、SwissADME及SwissTargetPrediction数据库筛选和收集药物有效成分对应的靶点。通过GeneCards数据库、OMIM数据库、TTD数据库、DrugBank数据库收集DKD的靶点。使用Cytoscape软件构建中药-成分-靶点网络图,STRING在线软件构建PPI网络,通过DAVID数据库进行交集靶点的基因本体(GO)功能及京都基因和基因组百科全书(KEGG)通路富集分析。根据网络药理学结果进行细胞实验验证。[结果] 筛选得到丹蛭降糖胶囊有效成分66种,涉及368个靶点,与DKD共同靶点54个,PPI分析显示,丹蛭降糖胶囊主要作用于IL-6、TP53、IL-1β、AKT1、MMP9、CCL2、TNF等靶点,共同靶点主要相关的生物学过程有365种,分子功能有63种,细胞组分有35种,作用在119条信号通路上发挥治疗DKD的作用,其中PI3K/Akt为关键信号通路。体外细胞实验证明,丹蛭降糖胶囊含药血清可降低PI3K/Akt信号通路相关蛋白的表达。[结论] 研究运用网络药理学预测了丹蛭降糖胶囊的核心靶点和通路并通过体外实验进行了相关验证,表明丹蛭降糖胶囊可通过多途径-多靶点-多通路对DKD发挥治疗作用。

Abstract:

[Objective] To explore the mechanism of action of Danzhi Jiangtang Capsule in intervening diabetic kidney disease(DKD) using network pharmacology combined with cellular experiments. [Methods] The active ingredients of Danzhi Jiangtang Capsule were collected through TCMSP and HERB databases,and the targets corresponding to the active ingredients of the drugs were screened and collected using PubChem database,SwissADME and SwissTargetPrediction databases. The targets of DKD were collected through GeneCards database,OMIM database,TTD database,and DrugBank database. Cytoscape software was used to construct the network map of traditional Chinese medicine(TCM)-components-targets,STRING online software was used to construct the PPI network,and DAVID database was used to analyze the GO function and KEGG pathway enrichment of the intersecting genes. Cellular experimental validation based on network pharmacology results. [Results] The screening obtained 66 kinds of active ingredients in Danzhi Jiangtang Capsule,involving 368 targets,and 54 common targets with DKD. PPI analysis showed that Danzhi Jiangtang Capsule mainly acted on the targets of interleukin-6(IL-6),Tumor Protein 53(TP53),interleukin-1β(IL-1β),protein kinase B 1(AKT1),matrix metalloproteinase(MMP),C-C motif chemokine ligand 2(CCL2),and tumor necrosis factor(TNF),and there were 365 kinds of biological processes related to common targets,63 kinds of molecular functions,and 35 kinds of cellular components,which acted on 119 signaling pathways to play a therapeutic effect for DKD,of which phosphoinositide 3-kinase(PI3K)/AKT was the key signaling pathway. In vitro cellular experiments demonstrated that Danzhi Jiangtang Capsule-containing serum decreased the expression of proteins related to the PI3K/AKT signaling pathway. [Conclusion] In this study,we used network pharmacology to predict the core targets and pathways of Danzhi Jiangtang Capsule and verified them through in vitro experiments,indicating that Danzhi Jiangtang Capsule can exert therapeutic effects on DKD through multiple pathways-multi-targets-multi-pathways.