基于YWHAZ/p38MAPK/CASP3信号通路探讨软脉煎治疗动脉粥样硬化的机制

Mechanism of Ruanmai decoction in treating atherosclerosis based on YWHAZ/p38MAPK/CASP3 signaling pathway

目的通过网络药理学、分子对接、动物实验探索中药复方软脉煎治疗动脉粥样硬化的作用机制。方法采用TCMSP查询软脉煎药物化学成分及靶点。在DrugBank、GeneCards、OMIM、TTD检索获得动脉粥样硬化的相关靶点。借助Cytoscape软件构建“药物-活性成分-靶点”PPI网络。利用David数据库进行GO及KEGG富集分析。采用Seesar软件将关键成分与核心靶点进行分子对接验证。动物实验使用高脂饲料喂养ApoE^(-/-)小鼠建立动脉粥样硬化小鼠模型,软脉煎颗粒剂灌胃,将主动脉病理切片染色,测定血脂,免疫荧光检测Mac2、YWHAZ蛋白表达,Western blot检测p-p38MAPK、c-CASP3蛋白表达。结果软脉煎共筛选了72个药物活性成分,对应168个治疗动脉粥样硬化的靶点基因。靶点主要富集在脂质代谢、炎症和免疫、氧化应激、血管内皮功能、细胞增殖与凋亡、糖酵解以及泛素化相关的生物过程及MAPK、TNF、PI3K-Akt、IL-17信号通路。动物实验验证了软脉煎可以通过下调关键靶点YWHAZ及p-p38MAPK、c-CASP3调控p38MAPK信号通路,减轻动脉粥样硬化炎症反应及炎症诱发的凋亡。结论软脉煎可以抑制YWHAZ表达及p38MAPK信号通路的激活,可能通过调控MAPK、TNF、PI3K-Akt、IL-17信号通路改善血管炎症、脂质代谢、氧化应激等病理过程,进而防治动脉粥样硬化。

Aim To explore the mechanism of action of Ruanmai decoction in treating atherosclerosis through network pharmacology.Methods The chemical components and targets of Ruanmai decoction were queried using TCMSP.Relevant targets for atherosclerosis were retrieved from DrugBank,GeneCards,OMIM,and TTD databases.The“Drug-Active Ingredient-Target”PPI network was constructed using Cytoscape software.GO and KEGG enrichment analysis were performed using the David database.Molecular docking verification of key components with core targets was conducted using the Seesar software.Atherosclerosis mouse models were established by feeding ApoE^(-/-)mice with a high-fat diet,and Ruanmai decoction granules were administered orally.Aortic pathological sections were stained,blood lipids were measured,and immunofluorescence was used to detect Mac2 and YWHAZ protein expression.Western blot was used to detect p-p38MAPK and c-CASP3 protein expression.Results Ruanmai decoction screened a total of 72 active drug components corresponding to 168 target genes for the treatment of atherosclerosis.The targets were primarily enriched in biological processes related to lipid metabolism,inflammation and immunity,oxidative stress,vascular endothelial function,cell proliferation and apoptosis,glycolysis,and ubiquitination.Signaling pathways such as MAPK,TNF,PI3K-Akt,and IL-17 were also involved.Animal experiments verified that RMJ could regulate the p38MAPK signaling pathway by down-regulating key targets YWHAZ,p-p38MAPK,and c-CASP3,thereby reducing AS inflammation and inflammation-induced apoptosis.Conclusions Ruanmai decoction can inhibit the expression of YWHAZ and activate the p38MAPK signaling pathway,potentially improving vascular inflammation,lipid metabolism,oxidative stress,and other pathological processes by regulating the MAPK,TNF,PI3K-Akt,and IL-17 signaling pathways,thus preventing and treating atherosclerosis.