基于网络药理学的泻心汤效应成分抗动脉粥样硬化作用机制研究

Mechanism of Xiexin Tang effective components on arresting atherosclerosis based upon network pharmacology

目的:基于网络药理学方法结合细胞实验研究泻心汤效应成分抗动脉粥样硬化(atherosclerosis,AS)的作用机制。方法:通过TCMSP等数据库获取泻心汤效应成分的靶点;同时采用GeneCards、TTD、Drugbank数据库检索AS相关的疾病靶点;运用Draw Venn Diagram软件构建泻心汤效应成分和AS的共同靶点;运用Cytoscape 3.7.2软件构建“效应成分-靶点”网络;利用STRING 11.0数据库构建共同靶点相互作用PPI网络;利用Metascape数据库进行GO功能及KEGG通路富集分析。最后,通过棕榈酸刺激EA.hy926内皮细胞建立体外炎症模型,对网络药理学的相关结果进行验证。结果:筛选得到泻心汤效应成分相关的靶点285个,疾病靶点589个,两者有交集靶点49个;网络拓扑分析得到核心靶点LDLR、RPPARG、VEGFA、IL-1β、I L-6、TNF-α等;KEGG分析得到PPAR、PI3K/AKT等信号通路,这些通路主要涉及炎症、脂质与动脉粥样硬化等相关途径,初步提示泻心汤效应成分可能通过作用于核心靶点调控PPAR、PI3K/AKT等信号通路以拮抗AS。ELISA和qPCR实验结果证明,泻心汤效应成分可通过降低炎症因子IL-1β、IL-6、TNF-α的分泌和mRNA的表达,抑制内皮细胞炎症反应,从而发挥抗AS作用。结论:泻心汤效应成分可通过作用于炎症因子IL-1β、IL-6、TNF-α等关键靶点发挥抗AS作用,这为泻心汤效应成分抗AS的机制研究提供了参考依据。

OBJECTIVE To explore the mechanism of Xiexin Tang effective components(XXT ECs)acting on atherosclerosis(AS)through a combination of network pharmacology and cell experiments.METHODS The targets of XXT ECs were obtained from the database of TCMSP while the disease targets of AS from the databases of GeneCards,TTD and Drugbank databases.The common targets of XXT ECs and AS were identified by the Venn Diagram software.The effective components-target network was constructed with Cytoscape 3.7.2 software.A common target interaction protein-protein interaction(PPI)network was created by the database of STRING 11.0.GO function and KEGG pathway enrichment analyses were performed with Metascape.An in vitro model was established by inducing EA.hy926 cells with palmitic acid(PA)for validating the results of network pharmacology.RESULTS The screening process yielded 285 corresponding protein targets,589 disease targets and 49 intersection targets.The core targets included LDLR,RPPARG,VEGFA,IL-1β,IL-6 and TNF-α.KEGG analysis primarily included pathways related to inflammation,lipid metabolism and endothelial protection.Enzyme-linked immunosorbent assay(ELISA)and quantitative polymerase chain reaction(qPCR)demonstrated that XXT ECs significantly suppressed inflammatory responses of palmitic acid-induced EA.hy926 endothelial cells leading to down-regulations of IL-1β,IL-6 and TNF-α.CONCLUSION XXT ECs may arrest atherosclerosis through targeting key inflammatory factors.The experiment provided valuable rationales for elucidating the anti-AS mechanism of XXT ECs.