不同运动能力小鼠在定制负荷运动适应中骨骼肌的基因转录响应——以细胞自噬与线粒体有关基因为例  

Transcriptional Responses of Skeletal Muscle to Customized Running Protocol in Mice with Different Exercise Capacity:Taking Autophagy and Mitochondrial-related Genes as Examples

作　　者：漆正堂[1,2] 刘静霞[1,3] 钱帅伟 王贵平[1,5] 邹勇 丁树哲[1,2] QI Zhengtang;LIU Jingxia;QIAN Shuaiwei(Key Lab of Adolescent Health Assessment and Exercise Intervention of Ministry of Education, East China Normal Univ., Shanghai 200241, China;School of P.E. and Health, East China Normal Univ., Shanghai 200241, China;Dept. of P.E., Southwest Jiaotong Univ., Chengdu 610031, China;School of Health Science, Wuhan Sports Univ., Wuhan 430079, China)

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 [2]华东师范大学体育与健康学院,上海200241 [3]西南交通大学体育工作部,四川成都610031 [4]武汉体育学院健康科学学院,湖北武汉430079 [5]苏州大学体育学院,江苏苏州215021

出　　处：《武汉体育学院学报》2022年第2期93-100,共8页Journal of Wuhan Sports University

基　　金：国家自然科学基金项目(31300977);中央高校基本科研业务费项目(40500-20104-222288)。

摘　　要：运动能力的差异是生物个体间客观存在的。本研究以细胞自噬与线粒体有关基因为例,探讨不同年龄不同运动能力小鼠在运动适应中骨骼肌进行基因选择性表达的转录调控机制,以及骨骼肌基因响应的个性化特征。将清洁级ICR小鼠分为青年对照组(YC)、青年运动组(YR)、成年对照组(AC)、成年运动组(AR),每组10只。采用递增负荷的运动能力测试确定青年、成年小鼠可以承受的最大跑速,YR、AR组小鼠按各自最大跑速的65%~75%进行耐力训练,每天训练1 h,持续4周。取腓肠肌测试mtDNA、ATP含量以及Caspases酶活性,实时荧光定量PCR检测mRNA表达,Western blot检测蛋白表达,用染色质免疫沉淀+PCR(ChIP-PCR)检测p53、ERRα与靶基因启动子结合的DNA片段。结果表明(1)骨骼肌mtDNA含量在成年小鼠运动后显著提高,ATP含量在成年、青年小鼠运动后均显著提高。(2)运动显著提高成年小鼠Caspase 3,8,9的酶活性,但是对青年小鼠无显著影响。(3)成年、青年小鼠自噬基因对运动响应显著,但线粒体生物发生、COX复合物、代谢调控有关基因只在成年小鼠对运动响应显著。(4)运动促进p53、ERRα与靶基因Tfam、SCO2、PUMA、Bax启动子的结合,但在成年、青年小鼠中靶基因转录水平不一致。结论即使保持相对一致的运动负荷,青年小鼠骨骼肌自噬与线粒体有关基因对运动的响应也比成年小鼠低。尽管运动促进p53、ERRα与靶基因启动子的结合,但靶基因在mRNA水平并不一定表达上调。Differences in exercise capacity exist among individuals.In this study,autophagy and mitochondria-related genes were taken to explore the transcriptional regulationof gene selective expression in skeletal muscle and the personalized characteristics of skeletal muscle gene response during exercise in mice with different ages and different exercise abilities.ICR mice were randomly divided into young control group(YC),young exercise group(YR),adult control group(AC)and adult exercise group(AR),with 10 mice in each group.The maximum running speed of each mouse was determined by increasing load exercise capacity test.Mice in YR and AR group were subjected to endurance training at 65%to 75%of their maximum running speed,for 1 hour a day for 4 weeks.Gastrocnemius was used to determine mtDNA content,ATP content and Caspases activity.Real-time PCR was used to detect mRNA levels.Western blot was used to detect protein content.DNA fragments binding to p53,ERRαwithin target gene promoters were detected by chromatin immunoprecipitation+PCR(CHIP-PCR).The results showed that:1)mtDNA content in skeletal muscle was significantly increased after exercise in adult mice,and ATP content was significantly increased after exercise in both adult and young mice.2)Exercise significantly increased the activity of Caspase 3,8 and 9 in adult mice,but had no significant effect on young mice.3)Autophagy-related genes were significantly responsive to exercise in adult and young mice,but the genes involved in mitochondrial biogenesis,COX complex and metabolic regulation were significantly responsive to exercise only in adult mice.4)Exercise promoted the binding of p53,ERRαto target genes Tfam,SCO2,PUMA and Bax promoters,but the transcription levels of target genes were inconsistent in adult and young mice.It could be concluded that the response of autophagy-and mitochondria-related genes in skeletal muscle to exercise in young mice was lower than that in adult mice,even with relatively consistent exercise load.Although exercise promoted the bi

关 键 词：运动 骨骼肌 基因选择性表达 染色质免疫沉淀 自噬 线粒体 

分 类 号：G804.2[文化科学—运动人体科学]