6周有氧运动对高脂膳食的ApoE敲除小鼠骨骼肌钙调控的影响  

Effects of 6-week aerobic exercise on calcium regulation in skeletal muscle of ApoE knockout mice fed by high-fat diet

作　　者：陈均 高扬 苏香楠 于亮[1,2] CHEN Jun;GAO Yang;SU Xiang-nan;YU Liang(School of Sport Science,Beijing Sport University,Beijing 100084;School of Strength and Conditioning Training Beijing Sport University,Beijing 100084,China)

机构地区：[1]北京体育大学运动人体科学学院,北京100084 [2]北京体育大学体能训练学院,北京100084

出　　处：《中国应用生理学杂志》2022年第2期137-142,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金(32071168);中央高校基本科研业务费专项资金资助(校2021ZD001)。

摘　　要：目的:探讨6周有氧运动对高脂膳食的载脂蛋白E(ApoE)基因敲除小鼠骨骼肌肌浆网钙调控蛋白的影响。方法:25只9周龄ApoE敲除小鼠(ApoE KO)随机选取5只ApoE KO小鼠进行最大跑速测试(以初始速度为4.8 m/min,坡度为0°,持续5 min后,每3 min速度增加1.2 m/min,直至力竭,最后速度为最大跑速,最大跑速的测试结果为(27.0±2.4)m/min,剩余20只ApoE KO小鼠随机分为ApoE KO小鼠高脂膳食组(KO)和ApoE KO小鼠高脂膳食+有氧运动组(KE),每组10只,同时以10只9周龄野生型C57BL/6J小鼠作为空白对照组(WT)。高脂饲料成分:脂肪含量为21%(w/w),胆固醇含量为1.5%(w/w)。KE组适应性训练1周后开始运动干预,运动方案为:40%最大跑速(10.8 m/min),运动时间40 min/d,频率每周3 d,共计6周。待末次运动后48 h,所有小鼠麻醉后经心脏穿刺处死后迅速分离双侧腓肠肌;可见光比色法检测骨骼肌Ca^(2+)浓度;Western blot法检测小鼠骨骼肌肌浆网钙调控蛋白RyR、CaM、CaMKⅡ、SERCA1、SERCA2蛋白表达。结果:与WT组相比,KO组小鼠骨骼肌Ca^(2+)浓度显著降低(P<0.01),骨骼肌肌浆网钙释放蛋白RyR、CaMKⅡ和钙回收蛋白SERCA1、SERCA2均显著降低(P<0.05),但CaM蛋白无显著变化;与KO组相比,KE组小鼠骨骼肌Ca^(2+)浓度和骨骼肌肌浆网钙回收蛋白SERCA1、SERCA2均显著升高(P<0.05),但骨骼肌肌浆网钙释放蛋白RyR、CaM、CaMKⅡ蛋白表达均无显著性差异。结论:高脂膳食可使ApoE敲除小鼠骨骼肌Ca^(2+)浓度降低、肌浆网钙释放作用和钙回收作用减弱,6周有氧运动训练能够显著提高其Ca^(2+)浓度、促进肌浆网钙回收作用。Objective:To investigate the effects of 6 weeks of aerobic exercise on the sarcoplasmic reticulum calcium regulatory proteins in skeletal muscle of apolipoprotein E(ApoE)knockout mice fed by high-fat diet.Methods:There were a total of twenty five 9-week-old ApoE knockout mice(ApoE knockout mice,ApoE KO),five of which were selected randomly for the maximum running speed test.The running speed would be increased by 1.2 m/min every 3 min after a 5-min duration of initial speed of 4.8 m/min without slope until exhaustion,then the final speed was set as maximal speed,and the test result of the maximum running speed was(27.0±2.4)m/min.The remaining 20 ApoE KO mice were randomly divided into ApoE KO mouse high-fat diet group(KO)and ApoE KO mouse high-fat diet+aerobic exercise group(KE),10 mice per group.Ten 9-week-old wild-type C57BL/6J mice were used as a blank control group(wild-type,WT).High fat diet composition:fat content was 21%(w/w)and cholesterol content was 1.5%(w/w).Exercise intervention was initiated 1 week after adaptive training in the KE group with an exercise protocol consisting of 40%maximal running speed(10.8 m/min),40 min/d and a frequency of 3 d/w for a total of 6 weeks.All mice were anesthetized and sacrificed by cardiac puncture then bilateral gastrocnemius muscles were isolated immediately 48 h after the final exercise.Skeletal muscle Ca^(2+) concentration was detected by visible light colorimetry;the expression levels of RyR,CaM,CaMKⅡ,SERCA1 and SERCA2,which are calcium regulated proteins of sarcoplasmic reticulum,in mouse skeletal muscle,was detected by Western blotting.Results:Compared with WT mice,the Ca^(2+) concentration,the expression levels of the sarcoplasmic reticulum calcium releasing proteins RyR,CaMKⅡ,and the calcium recycling proteins SERCA1 and SERCA2 were decreased significantly in skeletal muscle of the KO mice(P<0.05,P<0.01),while the expression of CaM protein did not change.Skeletal muscle Ca^(2+) concentration and the levels of sarcoplasmic reticulum calcium recycling prote

关 键 词：载脂蛋白E(ApoE) 有氧运动 骨骼肌 钙调控 小鼠 基因敲除 

分 类 号：Q445[生物学—生理学]