重复大强度运动对骨骼肌蛋白质组变异研究

    The Proteome Variation of Rat Skeletal Muscle Induced by Repeated Eccentric Exercise

    • 摘要: 目的:通过蛋白质组学研究手段分析重复大强度运动对骨骼肌损伤修复过程中蛋白质组表达情况,并从蛋白质组变异角度分析重复运动对骨骼肌损伤修复可能的作用机制;方法:72只Wistar大鼠随机分为正常对照组、一次离心运动组及一周后重复运动组。对照组不运动,其余组大鼠进行下坡跑,速度为18m/min,坡度-16°,运动时间为30min后休息5min,再运动30min。运动时间共60min。重复运动在一次运动后一周后进行。在运动后即刻、24h、48h、72h、168h取股四头肌进行蛋白质提取、双向电泳及特异蛋白质鉴定,并按照蛋白质功能对特异蛋白质进行分类;结果:重复运动后的修复过程中,能量代谢相关蛋白和细胞损伤修复相关蛋白表达在重复运动组中主要表现为下调为主,特别是在运动后0~72h之间。重复运动组中骨骼肌细胞结构蛋白0~24h内上调和下调数目无明显差异,在72h时,下调数目明显多于上调蛋白数目。损伤修复过程中表达变化的蛋白质中,能量代谢相关蛋白有异柠檬酸脱氢酶、磷酸葡萄糖变位酶、烟酰胺腺嘌呤二核苷酸脱氢酶、血色素结合蛋白、丙酮酸脱氢酶、磷酸丙糖异构酶,细胞损伤修复相关蛋白有泛素羧基末端水解酶、免疫球蛋白λ轻链、胞内氯离子通道蛋白、蛋白磷酸酶2C蛋白、谷胱苷肽过氧化物酶、双功能过氧化物酶等表达活跃,骨骼肌细胞结构蛋白有热休克蛋白、内质网驻地蛋白、角蛋白、肌球蛋白、线粒体内膜蛋白等。结论:重复运动后能量代谢关键酶表达,加速损伤细胞能量供应,同时在24h-48h之间进行可以减缓骨骼肌收缩蛋白降解、清除自由基、加快细胞胞吞、减缓炎症反应从而加快骨骼肌损伤修复。

       

      Abstract: Objective:To research proteome variety on skeletal muscle damage repair process induced by one and repeated eccentric exercise and proteomics mechanism of skeletal muscle repairing made by repeated eccentric exercise;Methods:72 Wistar rats were randomly divided into normal control group,one eccentric exercise group and repeat exercise group a week later.Control group was not done exercise,the rest groups were exercised with 18m/min of speed and-16 degrees of slope.Rats moved for 30min,rest for 5min and then moved for 30min.Repetitive motion was done after a week.Proteome extraction and dielectrophoresis were taken after 0h,24h,48h,72h,168h of one and repeated eccentric exercise,specific proteins were identified by MALDI-TOF MS and sorted according to its function.Results:Proteins expression about energy metabolism and cell damage were reduced especially from 0h to 72h after repeated motion.The proteins number of decreased and increased expression were not different obviously from 0h to 24h and the decreased proteins number was more than number of increased proteins obviously at 72h after repeated motion.During process of damage repair,energy metabolism related proteins included isocitrate dehydrogenase,phosphate glucose mutase,nicotinamide adenine dinucleotide dehydrogenase,hemoglobin binding protein,pyruvate dehydrogenase,triose phosphate isomerase;cell damage repair related proteins were consisted of ubiquitin carboxyl terminal hydrolytic enzyme,immune globulin lambda light chain,intracellular chlorine ion channel protein,protein phosphatase 2c protein,glutathione peroxidase,double function peroxidase and express active;skeletal muscle cell structure protein have heat shock protein,endoplasmic reticulum stations protein and keratin,myosin,mitochondrial membrane protein,etc.Conclusion:Repeated eccentric exercise may promote key enzyme expression of energy metabolism and energy supply for damage cell,accelerate skeletal muscle contraction protein degradation and cell swallow,and scavenge free radicals and slow inflammatory reaction during 2448h to speed up the skeletal muscle damage repair.

       

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