基于生物信息学和动物实验探讨苓桂术甘汤通过HIF-1α/HO-1信号通路改善心肌梗死后慢性心力衰竭的作用  

Bioinformatics and animal experiments reveal mechanism of Linggui Zhugan Decoction in ameliorating chronic heart failure after myocardial infarction via HIF-1α/HO-1 signaling pathway

作　　者：任涵 王舒舒 赵婉竹 徐少华 魏科东 吴婉婉 黄圣意 蔡瑞 张远红 黄金玲[1,3] REN Han;WANG Shu-shu;ZHAO Wan-zhu;XU Shao-hua;WEI Ke-dong;WU Wan-wan;HUANG Sheng-yi;CAIRui;ZHANG Yuan-hong;HUANG Jin-ling(School of Integrated Chinese Medicine and Western Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;School of Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;Anh ui Province Key Laboratory of Chinese Medicinal Formula,Hefei 230012,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽中医药大学中医学院,安徽合肥230012 [3]中药复方安徽省重点实验室,安徽合肥230012

出　　处：《中国中药杂志》2024年第23期6407-6416,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81973844,81373533)。

摘　　要：该研究旨在通过联合生物信息学和体内实验探究苓桂术甘汤(LGZGD)对心肌梗死(MI)诱导的慢性心力衰竭(CHF)小鼠心功能、心肌细胞自噬水平的影响,以及对缺氧诱导因子-1α(HIF-1α)/血红素加氧酶-1(HO-1)信号通路的调控作用。TCMSP数据库获取苓桂术甘汤活性成分及对应靶点,GEO、GeneCards和DisGeNET数据库收集疾病靶点;运用Cytoscape软件绘制“药物-成分-靶点”网络,在STRING网站进行蛋白-蛋白相互作用(PPI)网络分析,R语言进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析;分子对接验证核心靶点。采用手术结扎左前降支冠状动脉法构建MI后CHF小鼠模型,分为假手术组,模型组,苓桂术甘汤低、中、高剂量组(2.34、4.68、9.36 g·kg^(-1)),卡托普利组(3.25 mg·kg^(-1))。连续给药6周后使用多普勒超声成像系统检测小鼠心功能,包括左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)、左心室收缩末期内径(LVIDs)、左心室舒张末期内径(LVIDd);HE染色观察小鼠心肌组织形态学变化;RT-qPCR法检测小鼠心肌组织微管相关蛋白1轻链3B(LC3B)、自噬效应蛋白1(Beclin1)、自噬降解底物(p62)、HIF-1α、HO-1 mRNA表达水平;Western blot法检测小鼠心肌组织LC3B、Beclin1、p62、自噬相关蛋白5(ATG5)、HIF-1α、HO-1蛋白表达水平。网络药理学结果显示,共得到苓桂术甘汤有效成分103个,对应靶点224个,MI、CHF相关靶点分别为3 485、6 165个,GSE16499数据集获取差异基因3 263个,交集靶点共31个;核心药效成分为槲皮素、柚皮素和1-甲氧基菜豆素等;核心靶点为MAPK3、HMOX1(HO-1)、MYC、ADRB2、PPARD、HIF1A(HIF-1α)等;分子对接结果显示,核心靶点与苓桂术甘汤主要活性成分对接良好。苓桂术甘汤显著改善了MI后CHF小鼠的心功能,且心肌组织病理形态学变化明显减轻;Western blot和RT-qPCR结果显示模型组小鼠心肌组织中HIF-1α/HO-1信号通路和自噬过程This study aims to investigate the effect of Linggui Zhugan Decoction(LGZGD)on autophagy in the mouse model of chronic heart failure(CHF)induced by myocardial infarction(MI),as well as the regulatory effect of LGZGD on the hypoxia-inducible factor-1α(HIF-1α)/heme oxygenase-1(HO-1)signaling pathway,based on bioinformatics and animal experiments.The active ingredients and corresponding targets of LGZGD were retrieved from the Traditional Chinese Medicine Systems Pharmacology and Analysis Database,and GEO,GeneCards,and DisGeNET were searched for the disease targets.Cytoscape was used to establish a“drug-component-target”network.The protein-protein interaction(PPI)network analysis was performed on STRING.R language was used for gene ontology(GO)and Kyoto Encycloperfia of Genes and Genomes(KEGG)enrichment analyses.Molecular docking was adopted to validate the core targets.The mouse model of MI-induced CHF was established by surgical ligation of the left anterior descending coronary artery.The modeled mice were assigned into the sham,model,low-,medium-,and high-dose(2.34,4.68,and 9.36 g·kg^(-1),respectively)LGZGD,and captopril(3.25 mg·kg^(-1))groups.After continuous administration for 6 weeks,a Doppler ultrasound imaging system was used to examine the heart function indicators:left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-systolic dimension(LVIDs),and left ventricular end-diastolic dimension(LVIDd).The myocardial tissue was stained with hematoxylin-eosin for the observation of morphological changes.The mRNA levels of microtubule-associated protein 1 light chain 3 beta(LC3B),Beclin1,p62,HIF-1α,and HO-1 in the myocardial tissue were determined by RT-qPCR.The protein levels of LC3B,beclin1,p62,autophagy-related protein 5(ATG5),HIF-1α,and HO-1 were determined by Western blot.The results showed that 103 active components of LGZGD,corresponding to 224 targets,were obtained.A total of 3485 and 6165 targets related to MI and CHF,respectively,were retrieved.

关 键 词：心肌梗死后慢性心力衰竭 苓桂术甘汤 网络药理学 自噬 HIF-1α/HO-1信号通路 

分 类 号：R285.5[医药卫生—中药学]