基于网络药理学研究银丹心脑通软胶囊调控AKT抑制血管内膜增生的作用机制

Yindan xinnaotong capsule regulates AKT to inhibit neointimal hyperplasia:mechanism study based on network pharmacological analysis

目的采用网络药理学方法并结合体内实验验证,探讨银丹心脑通软胶囊(YDXNT)对血管内膜增生的干预效应及其机制。方法从TCMSP、Symmap数据库中检索YDXNT活性成分及其作用靶点,通过OMIM、DisGeNET、GeneCards数据库获取血管内膜增生的疾病靶点,并对YDXNT与血管内膜增生的靶点取交集。利用Cytoscape软件对交集基因构建“成分-疾病-靶点-通路”网络并可视化,应用Metascape进行GO和KEGG富集分析,CytoNCA插件获取核心靶点,利用autodock软件进行核心靶点与活性成分的分子对接。采用球囊损伤法制作大鼠血管内膜增生模型,给予YDXNT干预。通过苏木素-伊红(HE)染色分析各组血管形态学变化,免疫组织化学法检测内膜增生血管中核心靶蛋白的表达。结果网络药理学分析提示YDXNT可通过槲皮素、木犀草素、黄芩素等药理成分作用于血管内膜增生关键靶点AKT1(蛋白激酶B的一种亚基),HE结果显示YDXNT能有效抑制血管内膜增生,免疫组化检测结果显示模型组中AKT、p-AKT(Thr308)蛋白较假手术组表达增高,YDXNT给药组较模型组AKT、p-AKT(Thr308)蛋白表达下调。结论YDXNT可通过调控AKT表达及其磷酸化进而发挥抗血管内膜增生作用。

Objective To investigate the intervention effect of Yin Dan Xin Nao Tong Ruan Jiao Nang(YDXNT)on neointimal hyperplasia by integrating the strategy of network pharmacology and experiment in vivo.Methods The active constituents of YDXNT and their targets were retrieved from TCMSP and Symmap databases.Disease targets of neointimal hyperplasia were obtained from OMIM,DisGeNET and GeneCards databases,and the intersection of targets for YDXNT and neointimal hyperplasia was selected.Cytoscape software was used to construct and visualize the“component-disease-target-pathway”network for intersection genes.Metascape was used for GO and KEGG enrichment analysis,CytoNCA plug-in was used to obtain core targets,and autodock software was used for molecular docking between the core targets and active ingredients.The neointimal hyperplasia model was made by balloon injury method and YDXNT was give for intervention.Hematoxylin-eosin(HE)staining was used to analyze the changes of blood vessel morphology,and the expression of core target protein was detected by immunohistochemistry.Results The network pharmacological analysis suggested that YDXNT could act on AKT1,a key target of neointimal hyperplasia,through quercetin,luteotin,baicalein and other pharmacological components.HE results showed that YDXNT could effectively inhibit neointimal hyperplasia.Immunohistochemical results showed that the expression of AKT and p-AKT(Thr308)protein in the model group was higher than that in the sham operation group.The protein expression of AKT and p-AKT(Thr308)in YDXNT group was down-regulated compared with that in model group.Conclusion YDXNT may play an anti-neointimal hyperplasia role by regulating AKT expression and phosphorylation.