高住低练通过SIRT3调节心肌线粒体生物合成和自噬  

Living High-training Low Regulates Myocardial Mitochondrial Biogenesis and Mitophagy Through SIRT3

作　　者：丁晓青 马春伟 高炳宏[2] Ding Xiaoqing;Ma Chunwei;Gao Binghong(School of Kinesiology,Shanghai University of Sport,Shanghai 200438,China;School of Athletic Performance,Shanghai University of Sport,Shanghai 200438,China)

机构地区：[1]上海体育学院运动科学学院,上海200438 [2]上海体育学院竞技运动学院,上海200438

出　　处：《中国运动医学杂志》2023年第1期37-47,共11页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金项目(31771316);上海市人类运动能力开发与保障重点实验室项目(11DZ2261100)。

摘　　要：目的:观察高住低练对心肌线粒体生物合成及线粒体自噬的影响,探讨去乙酰化酶Sirtuin 3(SIRT3)是否在其中发挥关键作用。方法:将24只雄性C57BL/6小鼠随机分为对照组(C组)、高住低练组(E组)、SIRT3抑制组(T组),高住低练+SIRT3抑制组(TE)组,每组6只。C组正常饲养,不进行任何干预;E组白天在氧浓度为13.8%的低氧环境下居住8小时,晚上在常氧下进行跑台训练;T组腹腔注射SIRT3抑制剂3-TYP(50 mg/kg/2 d);TE组结合E组和T组的干预方法。高住低练干预为1次/天、6天/周,3-TYP干预为每两天注射1次、3次/周,共6周。干预结束后称量小鼠体重和心脏重量。采用HE染色观察心肌细胞状态,Masson染色观察心肌纤维化程度,Western blot检测心肌线粒体生物合成和线粒体自噬相关蛋白的表达水平,免疫组织化学观察SIRT3、过氧化物酶体增殖物激活受体γ共激活因子α(PGC-1α)、E3泛素连接酶(Parkin)的定位和表达情况。结果:与C组相比,E组的心肌细胞大小正常,排列有序,血管周围胶原纤维含量正常,SIRT3蛋白表达增强(P<0.01),线粒体生物合成蛋白磷酸化腺苷酸活化蛋白激酶(p-AMPK)、磷酸化p38丝裂原活化蛋白激酶(p-P38 MAPK)、PGC-1α、线粒体转录因子A(TFAM)蛋白表达水平上升,线粒体自噬蛋白PTEN诱导激酶1(Pink1)、Parkin、Nip3样蛋白X(NIX)蛋白表达水平上升(P<0.01)。与E组相比,TE组心肌细胞增大,排列拥挤,血管周围纤维含量增加,SIRT3蛋白表达水平下降(P<0.01),p-AMPK、p-P38 MAPK、PGC-1α、核呼吸因子1(NRF1)、TFAM蛋白表达水平下降,Pink1、Parkin、NIX、线粒体外膜受体B细胞淋巴瘤2家族蛋白/腺病毒E1B-19kDa结合蛋白3(Bnip3)蛋白表达水平下降(P<0.05,P<0.01)。结论:6周高住低练促进了心肌线粒体生物合成和线粒体自噬,SIRT3在低氧训练调控心肌线粒体生物合成和线粒体自噬中发挥关键作用,并参与低氧训练对整体心肌组织形态的保护过程。Objective To observe the effect of living high-training low on mitochondrial biogenesis and mitophagy,and whether Sirtuin 3(SIRT3)plays a key role in it.Methods Totally 24 male C57BL/6 mice were randomly divided into a normoxic control group(group C),a living high-training low group(group E),an SIRT3 inhibition group(group T),and a living high-training low+SIRT3 inhibition group(TE),each of 6.Group C was fed normally without any intervention.Group E lived in a low oxygen environment with oxygen concentration of 13.8%for 8 hours during the day,and conducted treadmill training in normal oxygen environment at night six times a week.Group T received intraperitoneal injection of SIRT3 inhibitor 3-TYP(50 mg/kg/2 d),3 times a week,while TE group underwent the invervention for group E and T at the same time.After 6-week intervention,all groups were weighed their heart and whole body.Hematoxylin-eosin and Masson staining were conducted to observe the status of myocardial cells and myocardial fibrosis,respectively.Meanwhile,the levels of mitochondrial biogenesis and mitophagy were observed using Western blotting,while the location and expression of SIRT3,peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)and Parkin were evaluated using immunohistochemistry.Results Compared with group C,myocardial cells of group E were normal in size and orderly arranged,with normal content of perivascular collagen fiber and enhanced expression of SIRT3(P<0.01),mitochondrial biogenesis proteins including phosphorylation-AMPactivated protein kinase(p-AMPK),phosphorylation-p38 mitogen-activated protein kinase(p-P38 MAPK),PGC-1αand mitochondrial transcription factor A(TFAM),as well as the mitophagy-related proteins like PTEN-induced kinase 1(Pink1),Parkin and Nip3-like protein X(NIX)(P<0.01).Compared with group E,myocardial cells in group TE were enlarged and crowded,with increased content of perivascular fibers,and decreased protein expression of SIRT3(P<0.01),p-AMPK,p-P38 MAPK,PGC-1α,the transcription factor nuclear respi

关 键 词：高住低练 心肌 线粒体生物合成 线粒体自噬 SIRT3 

分 类 号：G804.2[文化科学—运动人体科学] R54[文化科学—体育学]