联合液相色谱-质谱分析和网络药理学探讨益肾清利方治疗特发性膜性肾病的作用机制

Integrating LC-MS and Network Pharmacology Analysis to Explore the Mechanism of Yishenqingli Formula in Treating Idiopathic Membranous Nephropathy

目的联合液相色谱-质谱(Liquid chromatography-mass spectrometry,LC-MS)分析和网络药理学探讨益肾清利方治疗特发性膜性肾病(idiopathic membranous nephropathy,IMN)的潜在靶点。方法通过BATMAN-TCM数据库和LC-MS定性分析确定益肾清利方药物活性成分,通过网络药理学的方法预测益肾清利方治疗特发性膜性肾病的生物过程及机制通路,并对筛选出的核心靶点进行分子对接及体外实验验证。结果益肾清利方共筛选出15个活性成分,其作用靶点与特发性膜性肾病疾病靶点取交集后得到72个核心基因。GO富集分析结果显示包含凋亡信号通路的调控、白细胞迁移、肽基酪氨酸磷酸化等的参与;KEGG通路富集分析结果显示益肾清利方治疗特发性膜性肾病涉及TNF、PI3K/AKT、MAPK等细胞凋亡相关信号通路。体外实验表明,益肾清利方可通过调控PI3K/AKT通路减少足细胞凋亡。结论益肾清利方通过多靶点、多通路治疗特发性膜性肾病,对C5b-9诱导的足细胞亚溶破模型有抗凋亡作用,其作用机制可能与TNF、PI3K/AKT、MAPK信号通路有关。

Objective This study aimed to explore the potential targets of Yishenqingli Formula in treating idiopathic membranous nephropathy(IMN)using a combination of liquid chromatography-mass spectrometry(LC-MS)analysis and network pharmacology.Methods The active ingredients of the Yishen Qingli Formula were identified through the BATMAN-TCM database and LC-MS qualitative analysis.The biological processes and mechanism pathways of the Yishen Qingli Formula in treating IMN were predicted using network pharmacology,and molecular docking and in vitro,experiments were conducted to verify the selected core targets.The core targets were selected and validated through molecular docking and in vitro experiments.Results A total of 15 active ingredients were selected from the Yishen Qingli Formula,and 72 core genes were obtained by intersecting its target with the IMN disease target.GO enrichment analysis results showed that the regulation of apoptosis signaling pathway,white cell migration,peptide tyrosine phosphorylation,and so on were involved;The KEGG pathway enrichment analysis results showed that the treatment of IMN with Yishen Qingli Formula involves apoptosis-related signaling pathways such as TNF,PI3K/AKT,MAPK,etc.In vitro,experiments have shown that Yishen Qingli Formula can reduce podocyte apoptosis by regulating the PI3K/AKT pathway.Conclusion Yishen Qingli Formula is a treatment for idiopathic membranous nephropathy through multiple targets and pathways.It has an anti-apoptotic effect on the C5b-9 induced podocyte sub-lysis model,and its mechanism of action may be related to the TNF,PI3K/AKT,MAPK signaling pathways.