通脉养心丸治疗不同性别老年冠心病患者的分子机制研究

Molecular Mechanism of Tongmai Yangxin Pill in the Treatment of Elderly Patients with Coronary Heart Disease of Different Gender

背景冠心病是临床常见的心血管疾病,且心血管疾病风险存在性别差异。通脉养心丸已被广泛用于治疗冠心病,但其具体的作用机制仍不清楚。目的采用生物信息学方法分析通脉养心丸治疗不同性别老年冠心病患者的分子机制。方法2021年6—10月,从基因表达综合数据库(GEO)获取GSE142008数据集,将治疗前3例女性患者、3例男性患者的外周血单个核细胞样本作为对照组,治疗后3例女性患者、3例男性患者的外周血单个核细胞样本作为实验组。采用R 3.6.3软件及相关安装包进行数据预处理及差异基因的筛选,利用STRING数据库对差异基因制作蛋白质互作网络(PPI),将所得结果导入Cytoscape软件(3.8.2版本)中的MCODE、cytoHubba插件以筛选基因,再利用韦恩图在线工具将筛选出来的基因进行对比以获得关键基因。使用R 3.6.3软件及相关安装包进行GO功能富集分析和KEGG通路富集分析。结果数据预处理结果显示:各个样本中位数基本在一个水平线上,提示样本间归一化程度好。差异基因筛选结果显示:老年女性、男性冠心病患者治疗前后比较差异基因分别有796、1159个。PPI及关键基因选取结果显示:在老年女性冠心病患者中,最终选取了BTBD1、FBXL5、KBTBD7等10个关键基因;在老年男性冠心病患者中,最终选取了PJA2、LTN1、BTBD1等10个关键基因。在老年女性冠心病患者中,GO功能富集分析结果显示,关键基因主要富集于翻译后的蛋白质修饰等生物过程,SCF泛素连接酶复合物等细胞组成,泛素样蛋白转移酶活性等分子功能;KEGG通路富集分析结果显示,关键基因主要与泛素介导的蛋白水解、昼夜节律通路相关。在老年男性冠心病患者中,GO功能富集分析结果显示,关键基因主要富集于蛋白酶体介导的泛素依赖性蛋白质分解代谢过程等生物过程,泛素连接酶复合物等细胞组成,泛素蛋白转移酶活性等分子功能;KEGG通路富集分析结果显示,关键基因主要与泛素介导的蛋白水解通路相关。结论通脉养心丸治疗不同性别老年冠心病患者具有多靶点和多通路协调的特点,在老年女性患者中,其作用机制主要与泛素介导的蛋白质水解、细胞自噬、炎症反应、铁代谢、生物钟、激素调节等有关;在老年男性患者中,其作用机制主要与泛素介导的蛋白质水解、细胞自噬、DNA损伤修复等有关。

Background Coronary heart disease is a common clinical cardiovascular disease,and there are gender differences in the risk of cardiovascular disease.Tongmai Yangxin pill has been widely used in the treatment of coronary heart disease,but its specific mechanism of action is still unclear.Objective To analyze the molecular mechanism of Tongmai Yangxin pill in the treatment of elderly patients with coronary heart disease of different gender by bioinformatics methods.Methods The GSE142008 data set was loaded from the gene expression omnibus datasets(GEO)from June to October 2021.The peripheral blood mononuclear cell samples of 3 female patients and 3 male patients before treatment were used as the control group,and the peripheral blood mononuclear cell samples of 3 female patients and 3 male patients after treatment were used as the experimental group.R 3.6.3 software and related installation packages were used to data preprocessing and screen differential genes.The STRING database was used to create protein-protein interaction networks(PPI)for the differential genes,and the results were imported into the MCODE and cytoHubba plugins in Cytoscape software(version 3.8.2)to screen genes,then the Venn diagram online tool was used to compare the screened genes to obtain key genes.The GO functional enrichment analysis and KEGG pathway enrichment analysis were performed by R 3.6.3 software and related installation packages.Results The data preprocessing results showed that the median of each sample was basically on the same horizontal line,indicating that the degree of normalization between samples was good.The results of differential gene screening showed that there were 796 and 1159 differential genes in elderly women and men with coronary heart disease before and after treatment,respectively.The results of PPI and key gene selection showed that 10 key genes including BTBD1,FBXL5,KBTBD7 were finally selected in elderly female patients with coronary heart disease;10 key genes including PJA2,LTN1,BTBD1 were finally selected in elderly male patients with coronary heart disease.In elderly female patients with coronary heart disease,GO functional enrichment analysis showed that key genes were mainly enriched in biological processes such as post-translational protein modification,cellular composition such as SCF ubiquitin ligase complex,and molecular functions such as ubiquitin-like protein transferase activity;KEGG pathway enrichment analysis showed that key genes were mainly related to ubiquitin-mediated proteolysis and circadian rhythm pathways.In elderly male patients with coronary heart disease,GO functional enrichment analysis showed that key genes are mainly enriched in biological processes such as proteasome-mediated ubiquitin-dependent protein catabolism,cellular composition such as ubiquitin ligase complex,and molecular functions such as ubiquitin protein transferase activity;the results of KEGG pathway enrichment analysis showed that the key genes were mainly related to the ubiquitin-mediated proteolytic pathway.Conclusion Tongmai Yangxin pill has the characteristics of multi-target and multi-channel coordination in the treatment of elderly patients with coronary heart disease of different gender.In elderly female patients,its mechanism of action is mainly related to ubiquitin-mediated proteolysis,autophagy,inflammatory response,iron metabolism,circadian clock,and hormone regulation;in elderly male patients,its mechanism of action is mainly related to ubiquitin-mediated proteolysis,autophagy,and DNA damage repair.