循证研究与临床转化联合GEO数据库和网络药理学方法探讨芪参益气滴丸治疗心肌梗死的分子机制

Molecular mechanism of Qishen Yiqi Gutta Pills in treating myocardial infarction based on GEO database and network pharmacology

目的使用GEO数据库和网络药理学方法探讨芪参益气滴丸(QSYQ)干预心肌梗死(MI)的疗效基础和作用机制。方法GEO数据库筛选MI差异基因;TCMSP数据库以及文献检索筛选QSYQ的相关活性成分,SwissTargePrediction数据库预测成分的靶点;对差异基因和成分靶点取交集,Cytoscape软件构建“活性成分-交集靶点”调控网络;STRING数据库构建蛋白互作网络,Cytoscape中MCODE插件对网络进行分析,得到显著基因模块;clusterProfiler等R包对交集靶点进行GO功能和KEGG通路富集分析;AutoDock Vina进行成分-靶点的分子对接。CIBERSORT算法对GEO数据进行免疫浸润分析,并对关键基因表达量和免疫细胞含量进行Spearman相关性分析。结果共得到MI差异基因687个,活性成分靶点777个,交集靶点48个。PTGS2、MMP9、STAT3可能为QSYQ治疗MI的关键靶基因,分子对接模拟了与之对接最好的三个有效成分,分别为隐丹参酮、山奈酚、木犀草素。KEGG结果表明,交集靶点主要富集在NF-κB信号通路、TNF信号通路、流体剪切应力与动脉粥样硬化等通路上。关键基因与活化的肥大细胞、中性粒细胞、活化的自然杀伤细胞具有显著正相关(P<0.05),与未活化的CD4记忆性T细胞呈显著负相关(P<0.05)。结论QSYQ治疗心肌梗死是以多成分、多靶点、多通路的方式进行的,研究初步确定了QSYQ疗效的物质基础、关键靶标和通路,以期为后续研究提供参考。

Objective To discuss the curative effect and effect mechanism of Qishen Yiqi Gutta Pills on myocardial infarction(MI)by using GEO database and method of network pharmacology.Methods MI differential genes were screened from GEO database.The relevant active ingredients of Qishen Yiqi Gutta Pills were screened from TCMSP database and through literature searching.The ingredient targets were predicted from SwissTargePrediction database.The intersection of differential genes and ingredient targets was gained and a“active ingredient-intersection target”regulatory network was constructed by using Cytoscape software.A protein interaction network was constructed from STRING database,and was analyzed by using MCODE plugin in Cytoscape software to get significant gene modules.The intersection targets were given the enrichment analysis of GO function and KEGG pathway by using R package including ClusterProfiler.The molecular docking of ingredient-target was performed by using AutoDock Vina.The GEO data was given an immune infiltration analysis by using CIBERSORT algorithm,and key gene expression and immune cell content were given a Spearman correlation analysis.Results There were totally 687 MI differential genes,777 active ingredient targets and 48 intersection targets obtained.PTGS2,MMP9 and STAT3 were possibly the key target genes in MI treatment with Qishen Yiqi Gutta Pills.Molecular docking simulation showed that the best 3 active ingredients were cryptotanshinone,kaempferol and luteolin.KEGG results showed that intersection targets were mainly enriched in NF-κB signaling pathway,TNF signaling pathway,fluid shear stress and atherosclerosis pathway.The key genes were positively correlated to activated mast cells,neutrophils and activated natural killer cells(P<0.05),and negatively correlated to resting CD4 memory T cells(P<0.05).Conclusion Qishen Yiqi Gutta Pills can be used for treating MI through a multi-ingredient,multi-target and multi-pathway manner.The study preliminarily identified the material basis,key target and pathways of curative effect of Qishen Yiqi Gutta Pills,in order to provide reference for subsequent researches.