大负荷运动后不同时相下骨骼肌线粒体的分裂及其过程  

Fission and Dynamic Process of Skeletal Muscle Mitochondria in Different Time Phase after a Heavy Load Exercise

作　　者：白胜超 陈圣菊 李俊平[2] 尚画雨 高扬 王瑞元[2] BAI Shengchao;CHEN Shengju;LI Junping;SHANG Huayu;GAO Yang;WANG Ruiyuan(Dept.of PE,Nanjing University of Science and Technology,Nanjing 210094,China;School of Sport Science,Beijing Sport University,Beijing 100084,China;School of Sport Medicine and Health,Chengdu Sport University,Chengdu 610041,China)

机构地区：[1]南京理工大学体育部,江苏南京210094 [2]北京体育大学运动人体科学学院,北京100084 [3]成都体育学院运动医学与健康学院,四川成都610041

出　　处：《天津体育学院学报》2021年第1期89-95,共7页Journal of Tianjin University of Sport

基　　金：国家自然科学基金项目(项目编号:31471133)。

摘　　要：目的:探究一次性大负荷运动后,不同时相下骨骼肌线粒体的分裂情况,为运动后骨骼肌恢复提供理论指导。方法:雄性SD大鼠随机分为安静对照组(C)和单纯运动组(E),E组进行一次性下坡跑离心运动,之后按取材时间点分为0、2、6、12、24、48和72 h等时相组。使用透射电镜观察线粒体分裂现象,免疫荧光检测TOMM20/DRP1共定位,Western Blot测定线粒体中DRP1蛋白量和骨骼肌中FUNDC1表达量,免疫共沉淀检测FUNDC1与OPA1、CNX、DRP1等蛋白的相互作用。结果:透射电镜下线粒体出现片段化现象,TOMM20/DRP1共定位以及线粒体DRP1蛋白量明显升高(P<0.05),均在12 h达到峰值且显著高于安静组(P<0.05),FUNDC1未有显著变化(P>0.05),但与OPA1、CNX、DRP1等蛋白出现相互结合,并分别在安静状态、运动后2 h和1h先后达到峰值。结论:一次大负荷离心运动后,大鼠运动性骨骼肌损伤部位线粒体结构发生变化,出现分裂现象,分裂程度以及分裂蛋白DRP1的表达均呈时相性的动态变化,在运动后即刻逐渐加剧,12和24 h最为明显,48和72 h逐渐恢复,其分裂的过程为运动后蛋白FUNDC1与OPA1分离,转而与CNX结合,之后聚集DRP1到线粒体上导致分裂发生。Objective:To investigate the process of mitochondria fission of skeletal muscle in different phase after a bout heavy-load exercise on rats,and to provide theoretical guidance for the recovery of skeletal muscle function in multi-temporal periods after exercise training. Methods:Male Sprague-Dawley rats were randomly divided into control group(C,n=8)and exercise group(E,n=56). The group E was subjected to a single bout downhill eccentric exercise and further divided into groups of 0,2,6,12,24,48 and 72 h according to the time points of the materials taken after the exercise. The phenomenon of mitochondrial fission was observed by transmission electron miscroscope,the TOMM20/DRP1 was determined by immunoflurescence detection,the amount of DRP1 protein in mitochondria and FUNDC1 expression in skeletal muscle were measured in Western Blot,and the interactions of FUNDC1/OPA1,FUNDC1/CNX and FUNDC1/DRP1 were detected by immunoprecipitation. Results:The fragmentation of mitochondria appeared under transmission electron microscopy. The co-location of TOMM20/DRP1 and the amount of mitochondrial DRP1 protein increased significantly(P<0.05),both of them peaked at 12 h and significantly higher than control group(P<0.05). There was no significant change in FUNDC1(P>0.05),but it combined with OPA1,CNX and DRP1,and peaked at resting state,2 h and 12 h after exercise,respectively. Conclusions:A single bout heavy-load eccentric exercise can cause mitochondria fission in rats with exercise-induced skeletal muscle damage,and the dynamic changes is related to time phase. The mitochondrial fragmentation and DRP1 protein relative level were most severe at12~24 h after exercise,and then gradually recovered at 48~72 h after exercise. The fission process was mainly about the protein FUNDC1 was separated from OPA1,and then bound to CNX,and then DRP1 was aggregated onto the mitochondria to cause fission.

关 键 词：大负荷运动 骨骼肌 线粒体分裂 

分 类 号：G804.2[文化科学—运动人体科学]