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*

doi:10.12047/j.cjap.6221.2022.025

摘要/Abstract

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Ca²⁺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Ca²⁺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Ca²⁺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Ca²⁺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Ca²⁺浓度、促进肌浆网钙回收作用。

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

Abstract: 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 Ca²⁺ concentration was detected by visible light colorimetry; the expression levels of RyR, CaM, CaMK Ⅱ, 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 Ca²⁺ 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 Ca²⁺ concentration and the levels of sarcoplasmic reticulum calcium recycling proteins SERCA1 and SERCA2 were increased significantly (P＜0.05) in KE mice compared with KO mice, but there were no significant differences in the expressions of the sarcoplasmic reticulum calcium release proteins RyR, CaM and CaMK II. Conclusion: High fat diet can reduce the concentration of Ca²⁺in skeletal muscle and weaken the release and recovery of sarcoplasmic reticulum calcium in ApoE knockout mice. 6-week aerobic exercise training can significantly increase its Ca²⁺concentration and promote the recovery of sarcoplasmic reticulum calcium.

Key words: apolipoprotein E, aerobic exercise, skeletal muscle, calcium regulation, mice, gene knockout

中图分类号:

Q445

陈均, 高扬, 苏香楠, 于亮. 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^*[J]. 中国应用生理学杂志, 2022, 38(2): 137-142.

CHEN Jun, GAO Yang, SU Xiang-nan, YU Liang. Effects of 6-week aerobic exercise on calcium regulation in skeletal muscle of ApoE knockout mice fed by high-fat diet[J]. CJAP, 2022, 38(2): 137-142.

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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

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