基于分子对接和分子动态模拟探讨葛根素缓解大鼠心肌缺血再灌注损伤的作用机制  

The Effect of Puerarin in Alleviating Myocardial Ischemia-reperfusion Injury in Rats Based on Molecular Docking and Molecular Dynamics Simulation

作　　者：张春艳 曹晓璐[1] 刘嵩[1] 黄瑞松 李姝雅 王江友 Zhang Chunyan;Cao Xiaolu;Liu Song(Institute of Pharmaceutical Process,Hubei Province Key Laboratory of Occupational Hazard Identification and Control,School of Medicine,Wuhan University of Science and Technology,Wuhan 430065,China)

机构地区：[1]武汉科技大学医学院制药工艺研究所,职业危害识别与控制湖北省重点实验室,武汉430065 [2]武汉科技大学附属武汉亚洲心脏病医院心内科,武汉430022 [3]武汉科技大学附属武汉亚心总医院心内科,武汉430058

出　　处：《华中科技大学学报（医学版）》2024年第1期26-32,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81801307);武汉市卫生健康科研基金资助项目(No.WX20Q31);湖北省自然科学基金资助项目(No.2022CFB249)。

摘　　要：目的 基于分子对接和动态模拟,联合整体动物水平研究,揭示葛根素缓解心肌缺血再灌注损伤的具体作用机制。方法 采用分子对接联合分子动态模拟技术预测葛根素与细胞沉默调节蛋白1(SIRT1)的结合潜力;结扎SD大鼠左冠状动脉前降支构建心肌缺血再灌注模型,观察给予葛根素对心肌损伤的保护作用,观察抑制SIRT1表达后葛根素对心肌损伤的保护作用变化。2,3,5-氯化三苯基四氮唑(TTC)染色评估梗死面积;TUNEL联合免疫荧光检测心肌细胞凋亡率和SIRT1表达;透射电镜观察心肌超微结构;Western blot检测铁死亡相关蛋白表达。结果 分子对接结果表明葛根素和SIRT1能够形成稳定的复合物发挥药效;在构建心肌缺血再灌注模型的大鼠体内给予葛根素处理,能明显改善心肌损伤,上调SIRT1、溶质载体家族成员11(SLC7A11)、谷胱甘肽过氧化物酶4(GPX4),并下调铁反应元件结合蛋白2(IREB2)表达;一旦抑制SIRT1蛋白表达后,则葛根素的心肌保护作用被抑制。结论 分子对接和分子动态模拟技术能较好地预测葛根素与主要靶点SIRT1的作用潜力,葛根素通过激活SIRT1通路抑制铁死亡从而缓解心肌缺血再灌注损伤。Objective To investigate the effects of puerarin on myocardial ischemia-reperfusion injury and its mechanism.Methods Molecular docking and dynamics simulation were utilized to predict the binding potential of puerarin and SIRT1.A myocardial ischemia-reperfusion model was established in SD rats by ligating the anterior descending branch of the left coronary artery.The protective effect of puerarin on myocardial injury was observed,and the therapeutic effect of puerarin was compared after inhibition of SIRT1expression.The infarct volume was detected using 2,3,5-triphenyltetrazolium chloride(TTC)staining.The apoptosis rate and SIRT1expression of cardiomyocytes were detected by using TUNEL combined with immunofluorescence.Transmission electron microscope was used to observe the myocardial ultrastructure.Western blot was performed to detect the expression of ferroptosis-related proteins.Results Molecular docking studies confirmed the formation of stable complexes between puerarin and SIRT1.Puerarin treatment significantly increased myocardial ischemia-reperfusion injury through upregulation of SIRT1,SLC7A11and GPX4expression,and downregulation of IREB2expression in rats.The protective effect of puerarin on myocardium was abolished once SIRT1protein expression was inhibited.Conclusion Molecular docking and molecular dynamics simulation techniques can accurately predict the interaction of puerarin,and the main target SIRT1.Puerarin inhibits ferroptosis by activating SIRT1pathway,thereby alleviating myocardial ischemia-reperfusion injury.

关 键 词：分子对接 分子动态 葛根素 心肌缺血再灌注 SIRT1 

分 类 号：R542.22[医药卫生—心血管疾病]