基于网络药理学和实验验证探讨枳实薤白桂枝汤治疗心肌梗塞的作用机制

Mechanism of Zhishi Xiebai Guizhi Decoction on myocardial infarction based on network pharmacology and experimental validation

目的采用网络药理学方法及体外实验验证,对枳实薤白桂枝汤(Zhishi Xiebai Guizhi Decoction,ZXGD)治疗心肌梗塞(myocardial infarction,MI)的作用进行研究并对其机制进行探讨。方法从TCMSP数据库检索ZXGD的化学成分及其靶点。以“myocardial infarction”“MI”作为关键词检索GeneCards、OMIM和DisGeNET数据库,剔除重复项,得到MI的相关靶点,并对ZXGD与MI的靶点取交集。构建基于活性成分与疾病交集靶点的蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,利用DAVID数据库对交集靶点进行GO和KEGG通路富集分析,结合STRING数据库与Cytoscape 3.7.2软件将交集靶点进行可视化处理,构建“药材-成分-靶点-疾病”网络。采用结扎心脏冠状动脉左前降支建立MI小鼠模型,给予ZXGD进行干预,通过超声心动图检测、Western blot对预测结果进行实验验证。结果网络药理学分析显示,ZXGD可能通过槲皮素、柚皮素、β-谷甾醇、木犀草素等药理成分作用于TNF-α、IL-1β、IL-6、VEGFA、IL-10等靶点,参与TNF信号通路,从而治疗MI;动物实验发现,与模型组比较,ZXGD组小鼠左心室心功能、流出道血流等超声指标明显升高(P<0.05);以及梗死心肌组织中IL-1β、TNF-α、IL-6表达水平明显降低(P<0.05),IL-10表达水平明显升高(P<0.05)。结论ZXGD能够通过调控TNF-α、IL-1β、IL-6和IL-10等炎症因子保护心肌,从而治疗MI。

Aim To explore the effects of Zhishi Xiebai Guizhi Decoction(ZXGD)in the treatment of myocardial infarction(MI)using the network pharmacology method and verifying by in vivo experiments and to reveal the underlying mechanism.Methods The chemical components of ZXGD and related targets were retrieved from the TCMSP database.The targets of MI were searched from the GeneCards,OMIM and DisGeNET databases,with the keywords"myocardial infarction"and"MI",and removing duplicates.The intersection of ZXGD and MI targets were obtained,and a protein-protein interaction(PPI)network based on the intersection of active ingredients and disease targets was constructed.The DAVID database was used to conduct GO and KEGG pathway enrichment analysis on the intersection targets.Combined with STRING database and Cytoscape 3.7.2 software,the intersection targets were visualized as a"medicine-component-target-disease"network.The MI mouse model was established by ligation of the left anterior descending coronary artery of the heart.ZXGD was given once a day for 14 days.The cardioprotective effects of ZXGD were examined by ultrasound cardiogram and Western blot.Results The results of network pharmacology analysis showed that the pharmacological components of ZXGD such as quercetin,naringenin,β-sitosterol,and luteolin maybe work on the targets like TNF-α,IL-1β,IL-6,VEGFA,and IL-10.Animal experiments found that compared with the model group,ZXGD significantly increased the left ventricular cardiac function,outflow tract blood flow,and other ultrasound indexes of the mice(P<0.05).Moreover,the expression levels of IL-1β,TNF-αand IL-6 in myocardial infarction tissue were significantly down-regulated by ZXGD(P<0.05),while the expression level of IL-10 was significantly up-regulated(P<0.05).Conclusion ZXGD protects against MI and improves heart function by regulating inflammatory factors including TNF-α,IL-1β,IL-6,and IL-10.