基于网络药理学、分子对接探讨熊果酸治疗缺血性脑卒中的分子机制

Network pharmacology prediction and molecular docking-based strategy to discover the potential target and mechanism of ursolic acid against ischemic stroke

目的运用网络药理学、分子对接方法,挖掘熊果酸(ursolic acid,UA)治疗缺血性脑卒中的作用靶点及其潜在的分子机制。方法通过中药系统药理学数据库与分析平台及SwissTargetPrediction数据库对UA进行靶点预测,在Gene Cards数据库中获取缺血性脑卒中的靶点,并构建UA-缺血性脑卒中交叠靶点,通过String平台构建交叠靶点蛋白互作(protein-protein interaction,PPI)网络图,应用Cytoscape 3.9.1软件对PPI网络图进行拓扑属性分析,筛选出关键靶点后,进行基因本体论(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,最后进行靶点-UA分子对接进行验证。结果共筛选得到UA相关靶点122个和3509个缺血性脑卒中靶点;获得UA-缺血性脑卒中交叠靶点98个及19个关键靶点;其中白细胞介素-6(interleukin 6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)、白细胞介素-1β(interleukin 1β,IL-1β)、信号转导及转录激活因子3(signal transducers and activators of transcription,STAT3)、过氧化物酶体增殖激活受体γ(peroxisome proliferative activated receptor gamma,PPARG)是排名前5的核心靶点;KEGG通路富集分析结果显示关键靶点主要富集在血脂和动脉粥样硬化、TNF信号通路、白细胞介素17(interleukin 17,IL-17)信号通路、晚期糖基化终末产物-受体(advanced glycation end products-receptor for advanced glycation end products,AGE-RAGE)信号通路;GO功能富集分析显示关键靶点主要涉及转录因子活性、神经元凋亡过程的正向调节、炎症反应等过程;分子对接结果显示UA和排名前5的核心靶点可以稳定的结合,结合能均小于0 kcal/mol。结论UA可能通过调节IL-6、TNF、IL-1β、STAT3、PPARG影响血脂和动脉粥样硬化、TNF信号通路、IL-17信号通路、AGE-RAGE信号通路,发挥抗炎、神经保护等作用,以治疗缺血性脑卒中。

Objective To elucidate the targets and potential mechanism by which ursolic acid(UA)exerts its therapeutic effects in the treatment of ischemic stroke using network pharmacology and molecular docking analysis.Method The target genes of UA were screened from traditional Chinese medicines systems pharmacology database and analysis platform and SwissTargetPrediction,a web server for target prediction.Ischemic stroke-related genes were obtained using GeneCards database,and the intersection targets of UA and ischemic stroke were extracted.Intersection targets were imported into the STRING database to construct a protein-protein interaction(PPI)network.The core target genes were obtained using Cytoscape 3.9.1.Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed on the core target genes,followed by molecular docking of UA with these core genes.Result All 122 corresponding UA target genes and 3509 corresponding ischemic stroke target genes were screened out.98 intersection targets and 19 core genes between UA and ischemic stroke were identified.In PPI network,the top five proteins with the highest degree of connectivity were interleukin 6(IL-6),tumor necrosis factor(TNF),interleukin-1β(IL-1β),signal transducers and activators of transcription(STAT3)and peroxisome proliferative activated receptor gamma(PPARG).GO functional enrichment analysis showed that biological functions mainly involved transcription factor activity,positive regulation of neuronal apoptosis,inflammatory response and other functions.KEGG pathway enrichment revealed that lipid and atherosclerosis,TNF signalling pathway,interleukin 17(IL-17)signalling pathway advanced glycation end products-receptor for advanced glycation end products(AGE-RAGE)signalling pathway were mainly involved.The results of molecular docking suggested that UA were stable in binding with five core proteins,and the binding energy between UA and five core proteins were all less than 0 kcal/mol.Conclusion UA can exert anti-inflammatory and neuroprotective effects by regulating core targets such as IL-6,TNF,IL-1β,STAT3,PPARG,and participating in lipid and atherosclerosis,TNF signalling pathway,IL-17 signalling pathway,and AGE-RAGE signalling pathway,and it plays a therapeutic role in ischemic stroke.