线粒体自噬在容量负荷致心肌重构中的作用  

Role of PINK1-Mfn2-Parkin-Mediated Mitophagy in the Development of Myocardial Remodeling

作　　者：崔永春[1] 叶振 王小康 朱瑾彦 孟亮 袁卫民[1] 贾六军[1] 王佩合 周燕文 彭鹏 李巨波[1] 杨建中 岳广新 王欣[1] CUI Yong-chun;YE Zhen;WANG Xiao-kang;ZHU Jin-yan;MENG Liang;YUAN Wei-min;JIA Liu-jun;WANG Pei-he;ZHOU Yan-wen;PENG Peng;LI Ju-bo;YANG Jian-zhong;YUE Guang-xin;WANG Xin(Animal Experimental Center,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,State Key Laboratory of Cardiovascular Disease,Beijing Key Laboratory of Pre-clinical Research and Evaluation for Cardiovascular Implant Materials,Beijing,100037,China;Department of Pharmacy,Suqian First Hospital,Suqian Clinical College of Xuzhou Medical University,Affiliated Hospital of kangda College of Nanjing Medical University,Suqian 223800,China)

机构地区：[1]北京协和医学院动物实验中心,中国医学科学院阜外医院,心血管植入材料临床前研究评价北京市重点实验室,心血管疾病国家重点实验室,北京市100037 [2]江苏省人民医院宿迁分院宿迁市第一人民医院药学部,江苏省宿迁市223800

出　　处：《中国分子心脏病学杂志》2020年第6期3639-3643,共5页Molecular Cardiology of China

基　　金：北京市自然科学基金面上项目(7172181);心血管植入材料临床前研究评价北京市重点实验室主任基金(2018-PT2-ZR04);国家自然科学基金面上项目(81970387);北京市科委科技支撑项目(Z101107052210004);宿迁市自然科学基金面上项目(K201918);江苏省青年医学人才项目(QNRC2016485);宿迁市科技支撑计划项目(S201625)。

摘　　要：目的探索PINK1-Mfn2-Parkin介导的线粒体自噬在心肌结构重塑发生发展过程中的作用。方法首先利用野生型C57 BL/6小鼠40只,雌雄各半,体重20~30 g,通过腹主动脉-下腔静脉瘘(ACF)方法建立心肌结构重构模型,小鼠随机分为对照组(n=10)、术后2周组(n=10)、术后5周组(n=10)和术后15周组(n=10),观察不同组别线粒体自噬途径关键蛋白的表达水平变化规律。然后,利用野生型C57BL/6小鼠40只,体重和性别要求同上(分为假手术组、ACF模型组、ACF+阿霉素(AA,10 mg/kg/d)治疗组、ACF+环孢菌素A(CSA,1.5 mg/kg/d)治疗组,每组10只),并以Mfn2基因敲除C57BL/6小鼠ACF模型(n=10)为对照,探索线粒体自噬在心肌重构过程中的作用。结果与对照组比较,术后5周时模型组重构心肌中PINK1的稳定性增强、激活了下游Parkin、自噬体标志物LC3II表达量升高,而P62的表达水平逐渐下降,线粒体自噬增强。术后15周心脏功能明显下降,线粒体自噬相关蛋白表达量与代偿期相反,此时AA处理组小鼠与模型组比较,线粒体自噬关键蛋白的表达量明显升高,小鼠心肌重构情况明显好转,能量代谢恢复了脂肪酸β氧化为主的高效产能模式,延缓了其心力衰竭进程。而CSA处理组小鼠的线粒体自噬水平明显受到抑制,心肌代谢以糖酵解为主,心肌重构以及心脏功能下降更为显著。Mfn2基因敲除组小鼠心脏结构和功能变化进程与环孢菌素处理组相似。结论线粒体自噬参与心肌能量代谢重塑过程,而且对于容量负荷过载导致的心脏功能变化有重要影响。Objective To investigate the role of PINK1-Mfn2-Parkin-mediated mitochondrial autophagy in the development and progression of myocardiac remodeling.Methods First,40 wild-type C57 BL/6 mice,half male and female,weighing 20-30 g,were used to establish a model of myocardiac remodeling through the abdominal aorta-inferior vena cava fistula(ACF)method.The mice were randomly divided into a control group(n=10),2 weeks post operation(WPO,n=10),5 WPO(n=10)and 15 WPO(n=10).The time-course changes of the expression levels of key proteins in mitochondrial autophagy pathway were determined.Then,another 40 wild-type C57 BL/6 mice with the same weight and gender requirements were used and were randomly divided into sham group,ACF model group,ACF+doxorubicin(AA,10 mg/kg/d)group and ACF+cyclosporin A(CSA,1.5 mg/kg/d)group,10 mice per group.Ten Mfn2 gene knockout C57 BL/6 mouse ACF model were used as a control to explore the role of mitochondrial autophagy in cardiac remodeling process.Results Comparing with sham group,the stability of PINK1 was enhanced,downstream parkin and Mfn2 were activated,an increase in the levels of LC3-II/LC3-I(the conversion of LC3-I to LC3-II)was observed in model group(P<0.05).However,in 5 WPO group,the expression levels of above-mentioned key proteins in mitophagic pathway reduced significantly and heart function markedly decreased.Whereas,treatment with AA significantly up-regulated the expression of mitochondrial autophagic key proteins.Myocardiac remodeling was significantly improved.Energy metabolism restores high-efficiency productivity model based on fatty acid beta oxidation and the progression of heart failure was delayed significantly.In the CSAtreated group,mitophagy was inhibited,myocardial metabolism was dominated by glycolysis,and myocardial remodeling and cardiac function decreased more significantly.The status of heart structure and function in Mfn2 knockout mice were similar to those in CSA treated mice.Conclusions Mitochondrial autophagy is involved and plays an important role in the

关 键 词：心肌重构 线粒体 自噬 

分 类 号：R541.6[医药卫生—心血管疾病]