基于网络药理学和实验验证的桂枝甘草汤抗心肌缺血再灌注损伤的作用机制研究  

Mechanism of Guizhi Gancao Decoction against myocardial ischemia-reperfusion injury based on network pharmacology and experimental verification

作　　者：李连雨 陈林霖 刘萍 LI Lian-yu;CHEN Lin-lin;LIU Ping(Key Laboratory of Traditional Chinese Medicine Resources and Compound Prescription,Hubei University of Chinese Medicine,Wuhan 430065,China;School of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学中药资源与中药复方教育部重点实验室,湖北武汉430065 [2]湖北中医药大学基础医学院,湖北武汉430065

出　　处：《中国中药杂志》2024年第3期798-808,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30901937,81774186)。

摘　　要：基于网络药理学探讨桂枝甘草汤(Guizhi Gancao Decoction,GGD)对心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MI/RI)大鼠的保护作用及可能的药理机制。首先通过中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)、SwissTargetPrediction数据库和文献检索获取GGD抗MI/RI的化学成分及作用靶点,采用STRING数据库和Cytoscape 3.7.2软件对交集靶点进行蛋白-蛋白相互作用(protein-protein interaction,PPI)网络分析,对核心靶点进行基因本体(Gene Ontology,GO)和京都基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析,并构建“药物-核心成分-靶点-通路”网络,采用AutoDock Vina软件对主要活性成分与关键靶点进行分子对接验证。然后构建MI/RI大鼠模型,观察大鼠心肌梗死面积变化,苏木素-伊红(hematoxylin-eosin,HE)染色和透射电子显微镜(transmission electron microscope,TEM)检测心肌细胞病理和超微结构改变,蛋白免疫印迹(Western blot,WB)检测心肌组织相关蛋白表达。从GGD中筛选得到75个活性成分,对应318个治疗MI/RI靶点。PPI筛选得到肿瘤蛋白p53基因(tumor protein p53,TP53)、丝氨酸/苏氨酸激酶3(serine/threonine kinase 1,AKT1)、信号传导及转录激活蛋白(signal transducer and activator of transcription,STAT3)、非受体酪氨酸激酶(non-receptor tyrosine kinase,SRC)、丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、丝裂原活化蛋白激酶3(mitogen-activated protein kinase 3,MAPK3)、肿瘤坏死因子(tumor necrosis factor,TNF)等46个核心靶点。GO和KEGG富集分析得知,核心靶点主要影响细胞增殖和迁移、信号转导、细胞凋亡、转录过程,涉及癌症、糖尿病并发症中的高级糖基化终末产物-受体(advanced glycation end products-receptor,AGERAGE)、MAPK等信号通路。分子对接结果显示,GGD核心成分肉桂醛、儿茶素和甘草This study employed network pharmacology to investigate the effect of Guizhi Gancao Decoction(GGD)on myocardial ischemia-reperfusion injury(MI/RI)in rats and decipher the underlying mechanism.Firstly,the chemical components and targets of GGD against MI/RI were searched against the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),SwissTargetPrediction,and available articles.STRING and Cytoscape 3.7.2 were used to establish the protein-protein interaction(PPI)network for the common targets,and then Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were carried out for the core targets.The"drug-active component-target-pathway"network was built.Furthermore,molecular docking between key active components and targets was conducted in AutoDock Vina.Finally,the rat model of MI/RI was established,and the myocardial infarction area was measured.Hematoxylin-eosin(HE)staining and transmission electron microscopy(TEM)were employed to detect cardiomyocyte pathology and ultrastructural changes.Western blot was employed to determine the expression of related proteins in the myocardial tissue.A total of 75 chemical components of GGD were screened out,corresponding to 318 targets.The PPI network revealed 46 core targets such as tumor protein p53(TP53),serine/threonine kinase 1(AKT1),signal transducer and activator of transcription 3(STAT3),non-receptor tyrosine kinase(SRC),mitogen-activated protein kinase 1(MAPK1),MAPK3,and tumor necrosis factor(TNF).According to GO and KEGG enrichment analyses,the core targets mainly affected the cell proliferation and migration,signal transduction,apoptosis,and transcription,involving advanced glycation end products-receptor(AGE-RAGE),MAPK and other signaling pathways in cancers and diabetes complications.The molecular docking results showed that the core components of GGD,such as licochalcone A,(+)-catechin,and cinnamaldehyde,had strong binding activities with the core target proteins,such as MAPK1 and MAPK3.The

关 键 词：网络药理学 桂枝甘草汤 心肌缺血再灌注损伤 MAPK信号通路 

分 类 号：R285.5[医药卫生—中药学]