Journal Article
Randomized Controlled Trial, Veterinary
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[Effects of 6-week aerobic exercise on calcium regulation in skeletal muscle of ApoE knockout mice fed by high-fat diet].

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.

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