中药新药与临床药理2026，Vol.37Issue(6)：1100-1109,10.DOI:10.19378/j.issn.1003-9783.2026.06.012
调控铁死亡防治心肌缺血再灌注损伤的关键基因及潜在中药化合物筛选
Analysis and Validation of Key Genes and Potential Herbal Compounds Regulating Ferroptosis Against Myocardial Ischemia-Reperfusion Injury Based on Network Pharmacology
摘要
Abstract
Objective To screen key genes and potential herbal compounds regulating ferroptosis for the prevention and treatment of myocardial ischemia-reperfusion injury(MIRI)based on network pharmacology,molecular docking technology,and in vitro cellular experiments.Methods(1)Ferroptosis-related genes were downloaded from the FerrDb V2 database,and MIRI-related targets were obtained from the GeneCards and OMIM databases.The intersection of these gene sets was taken to identify potential targets regulating ferroptosis for ameliorating MIRI.The intersecting genes were imported into the STRING platform to construct a protein-protein interaction(PPI)network of potential targets and screen core targets.The HERB database was used for reverse screening of potential herbal compounds regulating ferroptosis for MIRI prevention and treatment.A target-compound network was established using Cytoscape 3.9.1 software to screen core compounds.Molecular docking simulations between core targets and core compounds were performed using AutoDock software to identify potential effective compounds.(2)HL-1 cardiomyocytes were injured by hypoxia/reoxygenation(H/R).Cells were treated with different concentrations of celastrol(0.25,0.5,1.0 μmol·L-1),different concentrations of dioscin(0.25,0.5,1.0 μmol·L-1),and different concentrations of ursolic acid(20,40,80 μmol·L-1).After 6 hours of hypoxia and 2 hours of reoxygenation,relevant indicators were assessed.Cell viability was detected using the CCK-8 method.Lactate dehydrogenase(LDH)content in the cell culture supernatant was measured by ELISA.Reactive oxygen species(ROS)content and mitochondrial membrane potential(∆Ψm)were detected using a live-cell imaging system.The expression levels of glutathione peroxidase 4(GPX4)and transferrin(TF)were detected by immunofluorescence staining.Results(1)A total of 128 common targets were identified as potential targets regulating ferroptosis for ameliorating MIRI.Core targets included TP53,IL6,HIF1A,IL1B,STAT3,ALB,JUN,mTOR,PTEN,and SRC.Through reverse screening using the HERB database,470 potential herbal compounds regulating ferroptosis for MIRI prevention and treatment were identified.Core compounds included resveratrol,curcumin,dioscin,ursolic acid,honokiol,and celastrol.Ursolic acid,celastrol,withaferin A,triptolide,and dioscin showed stable binding to the core targets.Based on the molecular docking results,three herbal compounds with high docking activity—celastrol,dioscin,and ursolic acid—were selected for in vitro experimental validation.(2)Compared with the normal group,the model group showed significantly decreased viability of HL-1 cardiomyocytes(P＜0.01),significantly increased LDH leakage(P＜0.01),significantly increased cellular ROS level(P＜0.01),significantly decreased ΔΨm(P＜0.01),significantly downregulated GPX4 protein expression(P＜0.01),and significantly upregulated TF protein expression(P＜0.01).Compared with the model group,the 0.25 μmol·L-1 celastrol group,the 0.25-1.0 μmol·L-1 dioscin groups,and the 80 μmol·L-1 ursolic acid group showed significantly increased viability of HL-1 cardiomyocytes(P＜0.05,P＜0.01);the 0.25-1.0 μmol·L-1 dioscin groups showed significantly decreased LDH leakage(P＜0.05,P＜0.01);the 1.0 μmol·L-1 dioscin group showed significantly decreased ROS level(P＜0.01),significantly increased ΔΨm(P＜0.01),significantly upregulated GPX4 protein expression(P＜0.01),and significantly downregulated TF protein expression(P＜0.01).Conclusion Celastrol,dioscin,and ursolic acid may serve as potential herbal compounds regulating ferroptosis for the prevention and treatment of MIRI.Dioscin ameliorates H/R-induced injury in HL-1 cardiomyocytes by regulating the expression of ferroptosis-related proteins GPX4 and TF,alleviating oxidative stress and mitochondrial damage,and inhibiting ferroptosis.关键词
心肌缺血再灌注损伤/铁死亡/网络药理学/薯蓣皂苷/雷公藤红素/熊果酸/氧化应激/HL-1心肌细胞Key words
myocardial ischemia-reperfusion injury/ferroptosis/network pharmacology/dioscin/celastrol/ursolic acid/oxidative stress/HL-1 cardiomyocytes分类
医药卫生引用本文复制引用
陈原原,郭浩,付建华,张会雨,郭帆,刘子馨,高佳明,曹策,胥淑娟,李兰兰,张晓晴..调控铁死亡防治心肌缺血再灌注损伤的关键基因及潜在中药化合物筛选[J].中药新药与临床药理,2026,37(6):1100-1109,10.基金项目
国家自然科学基金项目(82174219) （82174219）
中国中医科学院科技创新工程项目(CI2023C043YLL) （CI2023C043YLL）
国家中医药管理局中医药重点学科建设项目(zyyzdxk-2023231). （zyyzdxk-2023231）
