天香丹改善心肌缺血再灌注损伤作用机制的网络药理学和分子对接技术研究

Exploring the Mechanism of Tianxiang Pills in Improving Myocardial Ischemia-reperfusion Injury Based on Network Pharmacology and Molecular Docking

目的通过网络药理学和分子对接技术研究天香丹改善心肌缺血再灌注损伤的药效物质基础和分子作用机制。方法借助中药系统药理学数据库和分析平台(TCMSP)并查阅文献获得天香丹中4味中药的主要活性成分,并将与活性成分相联系的靶基因借助UniProt数据库翻译成相应基因。于PubChem数据库得到红景天、新塔花活性成分的详细信息,可以在TCMSP平台上找到该活性物质的靶基因,就使用TCMSP的结果,否则使用SwissTargetPrediction平台得到预测靶基因;利用GeneCards、OMIM、DISGENET和TTD数据库得到疾病相关靶点,对活性成分靶点与疾病靶点取交集后采用DAVID数据库进行基因本体(GO)功能以及京都基因与基因组百科全书(KEGG)富集分析;选择与本课题相关的通路cAMP、mTOR、核转录因子-κB(NF-κB)使用mapper工具构建KEGG通路图。最后通过AutoDock Vina软件对关键靶点和化合物进行分子对接。结果筛选出新塔花、红景天、丹参、降香针对心肌缺血再灌注损伤的128个活性成分和386个潜在作用靶点。蛋白质-蛋白质相互作用(PPI)网络拓扑分析发现白细胞介素-6(IL-6)、蛋白激酶B1(Akt1)、肿瘤坏死因子(TNF)的含量、白细胞介素-1β(IL-1β)等是天香丹治疗心肌缺血再灌注损伤的关键靶点。GO功能分析发现天香丹生物过程主要集中在RNA聚合酶Ⅱ启动子的转录正调控、基因表达的正向调控、凋亡过程的负调控、对外源刺激的反应等多个方面;KEGG信号富集分析结果显示主要涉及血脂与动脉粥样硬化、糖尿病并发症中的AGERAGE信号通路、TNF信号通路、细胞凋亡、IL-17信号通路、前列腺癌、HIF-1信号通路、cAMP信号通路、NF-κB信号通路、mTOR信号通路等。分子对接结果显示活性物质与靶点蛋白之间存在较强的结合力,其中槲皮素与TNF的结合力最强。结论天香丹治疗心肌缺血再灌注损伤具有多成分、多靶点、多通路相互协同的特点,为今后天香丹的临床应用提供了理论基础和参考。

Objective:To explore the pharmacological substance basis and molecular mechanism of Tianxiang Pills in improving myocardial ischemia-reperfusion injury through network pharmacology and molecular docking technology.Methods:Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TC⁃MSP)and related literature were consulted to obtain the main active ingredients of the four traditional Chinese medicines in Tianxiang Pills.The target genes associated with the active ingredients were translated into corre⁃sponding genes using the UniProt database.Detailed information on the active ingredients of Rhodiola rosea and Ziziphora bungeana from the Pubchem database was obtained.If the target gene of the active substance could be found on the TCMSP platform,the results of TCMSP were used;otherwise,the SwissTargetPrediction platform was used to predict the target gene.Disease-related targets were obtained from GeneCards,OMIM,DISGENET and TTD databases,and the DAD database was used to analyze the gene ontology(GO)function and Kyoto Encyclope⁃dia of Genes and Genomes(KEGG)after the intersection of active ingredient targets and disease targets.Finally,the key targets and compounds were molecularly docked by AutoDock Vina software.Results:A total of 128 active ingredients and 386 potential targets were identified for the effects of Ziziphora bungeana,Rhodiola rosea,Sal⁃via miltiorrhiza,and Dallbergia on myocardial ischemia-reperfusion injury.Protein protein interaction(PPI)net⁃work topology analysis revealed levels of interleukin-6(IL-6),protein kinase B1(Akt1),tumor necrosis factor(TNF),and interleukin-1β(IL-1β)were the key targets of Tianxiang Pills in treating myocardial ischemia-reper⁃fusion injury.GO functional analysis revealed that the biological processes of Tianxiang Pills mainly focused on positive regulation of RNA polymeraseⅡpromoter transcription,positive regulation of gene expression,negative regulation of apoptosis process,and response to external stimuli.KEGG signal enrichment analysis showed that it mainly involved blood lipids and atherosclerosis,AGE-RAGE signaling pathway,TNF signaling pathway,apoptosis,IL-17 signaling pathway,prostate cancer,HIF-1 signaling pathway,cAMP signaling pathway,NF-κB signaling pathway,mTOR signaling pathway and other signaling pathways.Molecular docking results showed a strong bind⁃ing affinity between the active substance and the target protein,with quercetin having the strongest binding affinity with TNF.Conclusion:The treatment of myocardial ischemia-reperfusion injury with Tianxiang Pills has the characteristics of multi-component,multi-target,and multi pathway synergy,providing a theoretical basis and refer⁃ence for the clinical application of Tianxiang Pills in the future.