基于网络药理学和细胞实验探究心脉康片治疗动脉粥样硬化心血管病的作用机制

To Explore Mechanism of Xinmaikang Tablets in Treatment of Atherosclerosis Cardiovascular Disease Based on Network Pharmacology and Cell Experiment

目的:运用网络药理学和细胞实验验证方法研究心脉康片治疗动脉粥样硬化心血管病的潜在药理学作用机制。方法:使用BATMAN-TCM数据库检索心脉康片的组成药物并筛选得到药物活性成分与潜在作用靶标;在GeneCards,人类在线孟德尔遗传数据库(OMIM)检索与动脉粥样硬化相关的疾病靶标;取药物与疾病靶标交集映射获得治疗靶标;将治疗靶标上传至STRING数据库进行蛋白质-蛋白质相互作用(PPI)网络构建;运用Cytoscape软件制作“药物-活性成分-治疗靶标”网络图,使用网络拓扑算法筛选关键作用靶标;使用DAVID软件进行基因本体(GO)和京都基因与基因组百科全书(KEGG)功能富集分析;使用体外细胞实验方法对药物治疗关键靶点作用进行验证。结果:检索并筛选获得心脉康片药物的活性成分共19个,潜在靶标132个,动脉粥样硬化疾病相关靶标基因4703个,获得交集靶标84个;筛选得到心脉康片治疗动脉粥样硬化疾病的关键治疗靶标3个,包括钙调蛋白1(CALM1),L-型电压依赖钙离子通道Α1C亚基(CACNA1C)和磷脂酰肌醇-3-激酶催化亚基α(PIK3CA);GO富集共得出313条生物学过程,89条分子功能和53条细胞组分;KEGG功能富集共得出40条通路,包括嘌呤代谢,肾素分泌,cGMP/PKG信号通路等。体外细胞实验结果验证心脉康片可上调CALM1,CACNA1C的表达,下调PIK3CA的表达,抑制炎症反应的活性,对动脉粥样硬化疾病起到治疗作用。结论:心脉康片治疗动脉粥样硬化心血管病可能通过白桦脂醇、甲基丁香酚等化合物,经过嘌呤代谢,肾素分泌,cGMP/PKG信号通路等途径,作用于CALM1,CACNA1C,PIK3CA等靶标而发挥作用。

Objective:To investigate the potential pharmacological mechanism of Xinmaikang tablets in the treatment of atherosclerosis cardiovascular disease by using network pharmacology and cell experimental validation.Method:The components of Xinmaikang tablets were searched by BATMAN-TCM database and the active ingredients and potential targets were screened.The atherosclerosis related disease targets were searched in GeneCards and online mendelian inheritance in man(OMIM)disease databases.The therapeutic targets were obtained by mapping the intersection of the tablets and disease targets.Therapeutic targets were uploaded to STRING database to construct protein-protein interaction(PPI)network.Cytoscape software was used to create a"drug-active component-therapeutic target"network map,and a network topology algorithm was used to screen key action targets.David software was used for gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)function enrichment analysis.The key targets of drug therapy were validated by in vitro cell assay.Result:A total of 19 active ingredients,132 potential targets and 4703 atherosclerotic disease-related target genes of Xinmaikang tablets were retrieved and screened,and 84 intersection targets were obtained.3 key therapeutic targets of Xinmaikang tablets in the treatment of atherosclerotic diseases were screened,including Calmodulin 1(CALM1),voltage-dependent L-type calcium channel subunit alpha-1C(CACNA1C)and Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform(PIK3CA).A total of 313 biological processes,89 molecular functions and 53 cell components were obtained by GO enrichment.A total of 40 pathways were obtained from KEGG functional enrichment,including purine metabolism,renin secretion,CGMP/PKG signaling pathway and so on.In vitro cell experiment results verified that Xinmaikang tablets can upregulate the expression of CALM1 and CACNA1C,down-regulate the expression of PIK3CA,so as to inhibit the activity of inflammatory response,and play a therapeutic role in atherosclerotic diseases.Conclusion:Xinmaikang tablets may treat atherosclerosis cardiovascular disease through betulin,methyleugenol and other compounds,through purine metabolism,renin secretion,cGMP/PKG signaling pathway and other pathways,which acts on CALM1,CACNA1C,PIK3CA and other targets.