补阳还五汤入脑成分治疗缺血性中风潜在机制的网络药理学分析与验证

Network Pharmacological Analysis and Verification of the Potential Mechanism of Buyang Huanwu Decoction in Treating Ischemic Stroke

目的:基于网络药理学研究补阳还五汤(buyang huanwu decoction,BYHWD)入脑成分治疗缺血性中风(ischemic stroke,IS)的潜在生物学机制,并通过实验对预测通路进行初步验证。方法:利用TCMSP、化学专业数据库和Swiss数据库筛选BYHWD的作用靶点,通过DiGSeE、OMIM、TTD和DRUGBANK数据库挖掘IS的相关靶点,将BYHWD的作用靶点与IS靶点相映射得到BYHWD作用于IS的潜在靶点,应用STRING数据库构建PPI网络图,借助Metascape对潜在靶点进行GO和KEGG富集分析,将实验动物随机分为对照组、模型组、BYHWD组和依达拉奉组,通过大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)术建立大鼠IS模型,造模7 d后采用Western blot技术对丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)/细胞外调节激酶(extracellular regulated protein kinases,Erk)通路进行验证。结果:获得BYHWD治疗IS的潜在靶点60个,主要富集到白细胞介素(interleukin,IL)-17信号通路、MAPK信号通路等途径。动物实验结果显示,与模型组比较,BYHWD组大鼠缺血侧脑皮质中磷酸化细胞外调节蛋白激酶(phosphorylation extracellular regulated protein kinases,pErk)表达明显降低(P<0.01)。结论:BYHWD治疗IS具有多成分、多靶点、多通路的特点,BYHWD治疗IS的潜在机制可能与抑制MAPK/Erk信号通路的活性相关。

Objective:To study the potential biological mechanism of the brain components of Buyang Huanwu Decoction(BYHWD)in the treatment of ischemic stroke(IS)based on network pharmacology,and verify the predictive pathway through experiments.Methods:The targets of BYHWD were predicted in the TCMSP,Chemistry,and Swiss databases.The targets of IS were searched by DiGSeE,OMIM,TTD,and DRUGBANK databases.The potential targets of BYHWD on IS were obtained by the intersection of compound targets and disease targets.The STRING database was used to construct a PPI network diagram.The Metascape database was used for GO and KEGG pathways enrichment analysis.Experimental animals were randomly divided into sham-operation group,model group,BYHWD group,and edaravone group.The IS model was established by middle cerebral artery occlusion(MCAO).After 7 days of surgery,the MAPK/Erk pathway was verified by the western blot experiment.Results:60 potential targets of BYHWD for IS treatment were obtained.KEGG was mainly enriched in the IL-17 signaling pathway,MAPK signaling pathway,and other pathways.The results of animal experiments showed that compared with the model group,the phosphorylation level of pErk(phosphorylation extracellular regulated protein kinases,pErk)in the ischemic cerebral cortex of the BYHWD group was significantly reduced(P<0.01).Conclusions:BYHWD has the characteristics of multiple components,multiple targets,and multiple pathways in the treatment of IS.The potential mechanism of BYHWD in the treatment of IS is probably related to the inhibition of the MAPK/Erk signaling pathway.