耐力运动对营养性肥胖小鼠骨骼肌细胞自噬及线粒体自噬的影响  

The Effect of Endurance Exercise on Autophagy and Mitophagy in the Skeletal Muscle of Mice with Alimentary Obesity

作　　者：崔迪[1] 邱守涛[1] 王海燕[1] 贺杰[1] 漆正堂[1] 孙易[1] 丁树哲[1] 

机构地区：[1]华东师范大学青少年健康评价与运动干预教育部重点实验室,上海200241

出　　处：《体育科学》2014年第12期63-71,共9页China Sport Science

基　　金：国家自然科学基金资助项目(31171142);青少年健康评价与运动干预教育部重点实验室建设项目(40500-541235-14203/004)

摘　　要：目的:探讨耐力运动对营养性肥胖小鼠骨骼肌细胞自噬及线粒体自噬的影响及其分子机制。方法:采用6周高脂膳食干预诱导生长期ICR雄性小鼠营养性肥胖模型,建模成功后随机分为高脂膳食对照组(HC,6只)、普通膳食干预组(HN,6只)及普通膳食联合耐力运动干预组(HNE,6只),另选造模对照ICR小鼠为普通膳食对照组(NC,6只),HC组小鼠持续喂养6周高脂膳食,NC、HN、HNE饲以普通膳食,HNE组小鼠进行6周跑台耐力训练,速度按周递增即15m/min、22m/min、27 m/min、31 m/min、35 m/min、35 m/min,40 min/次,6次/周。末次训练24h后处死小鼠并采样,应用JC-1探针法检测线粒体膜电位,RT-PCR及Western Blotting技术检测相关基因及蛋白表达情况。结果:1)6周高脂膳食可诱导小鼠体重、LEE’s指数及附睾脂肪百分比显著增加(P<0.05),BG、TG、TC、HDL-C、LDL-C含量显著增加(P<0.05);2)与HC比,HNE组骨骼肌MHCⅠ、Ⅱa、Ⅱc表达均显著增加(P<0.05),HN、HNE组MHCⅡb mRNA表达均下调;3)与NC比,HC组骨骼肌线粒体膜电位显著降低(P<0.05),HC、HN、HNE组小鼠骨骼肌mtDNA拷贝数量下降(P<0.05);4)与NC组比,HC组小鼠细胞自噬相关信号分子ATG 13、LC3 mRNA表达下调,P62 mRNA表达上调;HN、HNE组BECN1、ATG 13及LC3mRNA含量显著增加(P<0.05),HC组LC3Ⅱ/Ⅰ比值下降,HN、HNE组小鼠骨骼肌p62蛋白水平下降,HNE组LC3Ⅱ/Ⅰ比值上调;5)HC组NIX mRNA表达与NC组比下调,而在HN、HNE组含量增加;HNE组PARK2、PARL mRNA较HN组上调;HC组PINK1蛋白含量有所下调,而在HN及HNE组普遍上调(P<0.05)。结论:1)长期高脂膳食干预可诱导营养性肥胖发生,引起机体代谢紊乱、骨骼肌线粒体功能异常;2)耐力运动结合膳食改善可有效控制营养性肥胖小鼠体重并促进肌球蛋白的表达;3)耐力运动结合膳食改善可缓解营养性肥胖机体细胞自噬水平的下降,而对线粒体自噬的影响主要通过作用于PINK1/Parkin信号以调控骨骼肌�Objective：To determine the effect of endurance exercise on cellular autophagy and mi- tophagy in skeletal muscle of obese mice and explore the underlying molecular mechanism. Methods ： After the alimentary obesity ICR mice modle constrction, the obese mice were ran- domly divided into high-fat diet control group （HC, n= 6）, normal diet amelioration group （HN, n= 6） and normal diet combined with endurance exercise group （lINE, n = 6）. The mice of normal diet control group during the constretion of alimentary obesity model were assigned as the normal control group （NC, n= 6）. The mice in groups NC, HN and HNE were fed with nomal diet while the mice of group HC were fed with high fat diet . The mice of group HNE underwent treadmill running for 6 weeks, 45rain/d, 6 times per week. Results ： 1） Fiigh-fat diet for 6 week increased the body weight, Lee＇s index and epididymal fat percentage of mice, while a series of blood index such as BG, TG, TC, HDL-C and LDL-C increased sig- nificantly. 2） The relative contents of myosin heavy chain Ⅰ , Ⅱ a and Ⅱ c mRNA were upregulated in group HNE, compared with group EC. 3） The mitochondrial membrance potential of skeletal muscle in group HC was lower than that of group NC, while the mtDNA relative content in group HC, HN and FINE were lower than NC. 4） Compared to NC, the autophagy related genes ATG13 and LC3 mRNA decreased while P62 mRNA expression level increased; the BECN1, ATG13 and LC3 mRNA relative content of FIN and HNE were higher than thosein HC （P〈0.05） and the ratio of LC3 Ⅱ / Ⅰof HNE increased in skeletal muscle compared with HC. 5 ） The mitophagy related gene NIX mRNA expression was lower in HC compared to that of NC,whereas it accerlarated in group HN and HNE; in group HNE, the PARK2 and PARL mRNA relative content increased compared to HN; the protein content of PINK ldown- regulated in HC and up-regulated obviously in group HN and HNE（P〈0.05）. Conclusion： 1 ） Long-term high-fat-diet intervention can lead t

关 键 词：耐力运动 营养性肥胖 细胞自噬 线粒体自噬 PINK1/Parkin 

分 类 号：G804.7[文化科学—运动人体科学]