运动预适应促进GSH/Bcl-2合成及相互作用抑制阿霉素心脏损伤  

Exercise Preconditioning Promotes GSH/Bcl-2 Synthesis and Interaction to Inhibit Cardiac Injury by Doxorubicin

作　　者：王传志 张双双 郝月蓉 罗立杰 原阳 WANG Chuanzhi;ZHANG Shuangshuang;HAO Yuerong;LUO Lijie;YUAN Yang(School of Physical Education,Qingdao University,Qingdao 266071,China;School of Physical Education,Jining College,Jining 273155,China)

机构地区：[1]青岛大学体育学院,山东青岛266071 [2]济宁学院体育学院,山东济宁273155

出　　处：《体育科学》2023年第1期60-73,共14页China Sport Science

基　　金：国家自然科学基金(32000830);中国博士后科学基金第14批特别资助(2021T140356)。

摘　　要：目的:基于代谢组学分析,探究GSH与B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)相互作用在一次性运动预适应(exercise preconditioning,EP)抑制阿霉素(doxorubicin,DOX)治疗癌症产生心肌毒性中的作用机制。方法:40只19周龄雌性SD大鼠随机分为对照(control,C)组、DOX组、EP组、EP+DOX组,每组10只。DOX组注射DOX,EP组大鼠进行1次大强度间歇运动,EP+DOX组于EP后24 h注射DOX。通过心脏超声检测大鼠心功能;通过化学荧光免疫分析法检测心肌损伤标志物表达水平;通过组织学染色、透射电镜观察和分析大鼠心肌组织病理变化;通过Western Blot检测心肌凋亡及促存活基因相关蛋白表达;通过代谢组学分析探究心肌代谢物和代谢通路变化;通过免疫共沉淀法检测Bcl-2与GSH转运蛋白2-酮戊二酸载体(2-oxoglutarate carrier,OGC)的结合活性;通过免疫荧光双标法分析GSH与Bcl-2共定位水平。结果:DOX组大鼠心肌线粒体肿胀、外膜破裂、体积增大、线粒体嵴排列紊乱甚至断裂,线粒体基质内出现云絮状结构。与C组相比,DOX组大鼠心功能显著更低;心肌损伤指标血清c Tn I、LDH、丙二醛含量显著更高;心肌细胞损伤、纤维化、炎症症状明显;心肌细胞抗凋亡蛋白Bcl-2表达水平显著更低,Bad、Bax、Caspase3等凋亡蛋白表达显著更高;YAP磷酸化水平显著更高;GSH、NAD+代谢水平显著更低;Bcl-2与OGC结合程度显著更低;Bcl-2与GSH共定位水平显著更低。与DOX组相比,EP+DOX组上述指标均呈不同程度改善。结论:经典形式的EP通过促进GSH/Bcl-2相互作用,可以减轻DOX治疗引起的心肌细胞凋亡及氧化应激损伤,抑制炎症及纤维化,改善心功能。Objective:To investigate the mechanisms of GSH interaction with Bcl-2 following one-time exercise precondition(EP)in reducing cardiotoxicity induced by DOX treatment of cancer.Methods:Forty 19-week-old female SD rats were randomly divided into control group(C),doxorubicin group(DOX),exercise preconditioning group(EP),and exercise preconditioning+doxorubicin group(EP+DOX),10 rats in each group.The Dox group rats were injected with doxorubicin,EP group rats were conducted about of high-intensity intermittent exercise,and the EP+DOX group rats were injected with doxorubicin 24 h after EP.Cardiac ultrasound was used to detect cardiac function in rats;chemical fluorescence immunoassay was used to detect the expression level of myocardial injury markers;histological staining and transmission electron microscopy were used to observe and analyze the histopathological changes of myocardium;Western blot was used to detect myocardial apoptosis and pro-survival gene-related protein expression;Metabolomics analysis was performed to explore the changes of myocardial metabolites and metabolic pathways;immunoprecipitation was used to detect the interaction between Bcl-2 and OGC(GSH transporter protein);immunofluorescence double-labeling was used to analyze the co-localization of GSH and Bcl-2.Results:In the DOX group,the myocardial mitochondria were swollen,the outer membrane was ruptured,the volume increased,and the mitochondrial cristae were disorganized or even broken;in the mitochondrial matrix,the linear dense cristae were visible,and the cloudy flocculent structures were appeared.Compared with group C,the cardiac function of rats in the DOX group was significantly decreased;the serum c Tn I,LDH,and MDA contents of myocardial injury indexes were significantly higher;the symptoms of myocardial cell injury,fibrosis,and inflammation were obvious;the expression levels of anti-apoptotic protein Bcl-2 and cardiomyocytes were significantly lower,and the expression of apoptotic proteins such as Bad,Bax and Caspase3 were significant

关 键 词：运动预适应 阿霉素 心脏毒性 B细胞淋巴瘤-2 谷胱甘肽 

分 类 号：G804.7[文化科学—运动人体科学]