AP39通过上调PINK1/Parkin介导的线粒体自噬改善尿毒症大鼠心肌纤维化  

AP39 ameliorates uremic myocardial fibrosis in rats by upregulating PINK1/Parkin-mediated mitochondrial autophagy

作　　者：宋熊 肖婷 刘达 聂连桂 刘茂军[1] 王森 赵俊雄 汪刘洋 刘星[1] 江振涛 杨军[1] SONG Xiong;XIAO Ting;LIU Da;NIE Liangui;LIU Maojun;WANG Sen;ZHAO Junxiong;WANG Liuyang;LIU Xing;JIANG Zhentao;YANG Jun(Department of Cardiology,First Affiliated Hospital,University of South China,Hengyang,Hunan Province,421001;Department of Cardiology,Shenzhen Longhua District Central Hospital,Shenzhen,Guangdong Province,518110;Department of Cardiology,Nanhua Hospital Affiliated to University of South China,Hengyang,Hunan Province,421001,China)

机构地区：[1]南华大学附属第一医院心血管内科,湖南衡阳421001 [2]深圳市龙华区中心医院心血管内科,广东深圳518110 [3]南华大学附属南华医院心血管内科,湖南衡阳421001

出　　处：《第三军医大学学报》2021年第24期2633-2639,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81870230);深圳市龙华区医疗卫生机构2020年区级科研项目(2020035);湖南省自然科学基金青年项目(2020JJ5505)。

摘　　要：目的阐明AP39对尿毒症大鼠心肌纤维化的改善作用及机制。方法取40只SD雄性大鼠按随机数字表法分成4组(n=10):假手术组(Sham组)、尿毒症心肌病模型组(UCM组)、AP39干预组(UCM+AP39)组及AP39对照组。除Sham组及AP39对照组外,另外两组大鼠采用5/6肾切除的经典手术方法建立尿毒症心肌病大鼠模型。术后UCM+AP39组及AP39对照组大鼠予以AP39 (100 nmol/kg 1次/d)腹腔注射,Sham组及UCM组大鼠予以同等体积生理盐水腹腔注射。超声成像平台检测各组大鼠左心室缩短分数(left ventricular fractional shortening,LVFS)的变化,Masson染色观察各组大鼠心肌间质组织中胶原沉积量,免疫组化观察各组大鼠心肌组织中胶原蛋白Ⅲ(CollagenⅢ,ColⅢ)蛋白量的表达,Western blot检测PINK1、Parkin、Atg5、LC3Ⅱ/Ⅰ、Beclin1和P62(也被称为SQSTM1)等蛋白在各组大鼠心肌组织的表达水平。结果与Sham组相比,UCM组中大鼠的LVFS下降,心肌胶原沉积量明显增多(P<0.05),ColⅢ表达增多(P<0.05),心肌组织排列紊乱,并且PINK1、Parkin、Atg5、LC3Ⅱ/Ⅰ、Beclin1等蛋白的表达量明显下调(P<0.05),P62蛋白表达明显上调(P<0.05);而与UCM组相比,UCM+AP39组大鼠的LVFS升高,心肌胶原沉积量明显减少(P<0.05),ColⅢ表达减少(P<0.05),并且PINK1、Parkin、Atg5、LC3Ⅱ/Ⅰ和Beclin1蛋白的表达量明显上调,P62蛋白表达明显下调(P<0.05)。结论 AP39能改善尿毒症大鼠心肌纤维化,其机制可能与上调PINK1/Parkin介导的线粒体自噬有关。Objective To elucidate the ameliorative effect of AP39 on myocardial fibrosis in uremic rats and investigate its possible mechanism.Methods Forty SD male rats were randomly and equally divided into 4 groups:Sham group,uremic cardiomyopathy group(UCM group),AP39 intervention group(UCM+AP39 group) and AP39 control group(n=10).The rats in the UCM group and the UCM+AP39 group were treated with classical operation of 5/6 nephrectomy to establish the uremic cardiomyopathy model,and then the rats in the latter 2 groups were given intraperitoneal injection of AP39(100 nmol/kg,once a day) for 4 weeks,while those of the Sham and model groups were intraperitoneally injected with the same amount of normal saline.Ultrasonic imaging platform was used to detect the value of left ventricular fractional shortening(LVFS) of rats in each group.Masson staining and immunohistochemical staining were performed respectively to observe collagen deposition and the expression of Collagen Ⅲ protein in the myocardial interstitial tissue of each group.Western blotting was adopted to determine the expression levels of PETN-induced putative kinase-1(PINK1),Parkinson’s disease protein(Parkin),autophagy-related gene-5(Atg5),microtubule-associated protein-1 light chain-3 Ⅱ/Ⅰ(LCEⅡ/Ⅰ),Beclin1 and P62(also called sequestosome,SQSTM1) in the myocardial tissue of each group.Results As compared with those in the Sham group,the rats in the UCM group had significantly decreased LVFS,increased collagen deposition content and Collagen Ⅲ protein level(P<0.05),and disordered arrangement in myocardial tissue.The expression levels of PINK1,Parkin,Atg5,LC3Ⅱ/Ⅰand Beclin1 were significantly reduced,while the level of P62 was enhanced in the UCM group(P<0.05).By contrast,the rats in the UCM+AP39 group had higher LVFS,lower collagen deposition content and Collagen Ⅲ protein level than those in the UCM group(P<0.05),the expression levels of PINK1,Parkin,Atg5,LC3 Ⅱ/Ⅰ and Beclin1 were remarkably up-regulated,and the level of P62 down-regulated(P

关 键 词：AP39 心肌纤维化 尿毒症心肌病 线粒体自噬 PINK1/Parkin通路 

分 类 号：R542.23[医药卫生—心血管疾病] R692.5[医药卫生—内科学]