黄连-川芎治疗冠状动脉粥样硬化和急性心肌梗死作用机制差异性的网络药理学分析

The Mechanism of Huanglian and Chuanxiong in the Treatment of Coronary Atherosclerosis and Acute Myocardial Infarction Based on Network Pharmacology

目的:基于网络药理学方法与分子对接技术,探究活血解毒中药药对黄连-川芎治疗冠状动脉粥样硬化(CAS)和急性心肌梗死(AMI)作用机制的差异性,为活血解毒中药在冠心病不同阶段的应用提供理论依据。方法:通过基因表达综合数据库(GEO)获得CAS和AMI的差异基因,并进行GEO差异分析。从中药系统药理学数据库与分析平台(TCMSP)搜集黄连、川芎中主要活性成分和筛选预测靶点。根据药物作用靶点,构建蛋白质-蛋白质相互作用(PPI)网络图并进行拓扑分析,再利用DAVID数据库进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)富集分析。采用Schrodinger软件将中药主要活性成分分别与CAS和AMI的关键蛋白进行分子对接。结果:黄连-川芎治疗CAS和AMI的活性成分基本相同。PPI网络拓扑分析显示,治疗CAS有17种靶蛋白,治疗AMI有37种靶蛋白,两者共有16个相同的靶蛋白,而AMI多出了21个特有靶蛋白等,表明在AMI时,黄连-川芎能在调控CAS相关靶点基础上进一步作用于新的靶点。GO富集分析显示黄连-川芎通过不同的生物过程干预CAS和AMI。KEGG富集分析发现CAS相关通路为22条,而AMI相关通路为90条,两者共有通路17条,但在CAS和AMI中的富集程度存在差异;而AMI特有的通路有73条,主要为肿瘤坏死因子(TNF)信号通路、核转录因子-κB(NF-κB)信号通路等,涉及多个炎症相关通路;表明中药黄连和川芎能够在调控CAS相关通路的基础上,进一步调控AMI时的特有通路。分子对接结果显示,主要化合物和靶点能很好地结合,支持网络药理学结果。结论:本研究阐释了黄连-川芎基于多化合物、多靶点、多途径治疗CAS和AMI的可能作用机制及两者的差异性,发现黄连-川芎可以在调控CAS和AMI共有靶点和通路的基础上,进一步作用于AMI发生时出现的特有蛋白和通路,揭示了在冠心病不同阶段活血解毒中药黄连-川芎作用的不同机制,从药理学层面完善了冠心病的“瘀毒”理论。

Objective:To investigate the difference of the mechanisms of Huanglian and Chuanxiong in the treatment of coronary atherosclerosis(CAS)and acute myocardial infarction(AMI)based on a network pharmacology approach and molecular docking technology.Methods:The differential genes of CAS and AMI were obtained through the comprehensive gene expression database(GEO).The main active ingredients and predicted targets in Huanglian and Chuanxiong were collected from the traditional Chinese medicine systematic pharmacology database and analysis platform(TCMSP).According to drug action targets,protein-protein interaction network diagram(PPI)was constructed and topological analysis was performed.David database was used for gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Schrodinger software was used to dock the key active ingredients of traditional Chinese medicine with the key proteins of CAS and AMI respectively.Results:The active ingredients of Huanglian and Chuanxiong in the treatment of CAS and AMI were similar.PPI network topology analysis showed that there were 17 target proteins for the treatment of CAS and 37 target proteins for the treatment of AMI,with a total of 16 identical target proteins,while AMI had 21 more specific target proteins,Huanglian and Chuanxiong could further act on new targets on the basis of regulating CAS related targets in AMI.GO enrichment analysis showed that blood activating and detoxifying herbs interfered with CAS and AMI through different biological processes.KEGG enrichment analysis showed that there were 22 CAS-related pathways and 90 AMI-related pathways,and there were 17 common pathways between CAS and AMI,but there were differences in the enrichment degree between CAS and AMI.There were 73 specific pathways of AMI,mainly tumor necrosis factor(TNF)signaling pathway,nuclear transcription factor-κB(NF-κB)signaling pathway,etc.,involving multiple inflammatory pathways.These results indicated that Huanglian and Chuanxiong could further regulate the specific pathway of AMI on the basis of the regulation of CAS related pathway.Molecular docking results showed that the main compounds and targets were well combined,supporting the network pharmacological results.Conclusion:In this study,the possible mechanism of action of Huanglian and Chuanxiong in the treatment of CAS and AMI based on multi-compound,multi-target and multi-pathway were explained.It was found that Chinese herbs could further act on the specific proteins and pathways in the occurrence of AMI on the basis of regulating the common targets and pathways of CAS.The different mechanisms of activating blood and detoxification therapy in different stages of coronary heart disease were revealed,and the theory of"stasis poison"of coronary heart disease was perfected from the pharmacological level.