Mechanism by which dysferlin promotes repair of exercise-induced skeletal muscie injury in rats

doi:10.3969/j.issn.2095-4344.0219

Abstract

Abstract:

BACKGROUND: Numerous studies concerning dysferlin focus on muscle diseases (such as muscular dystrophy), but the relationship between membrane repair after exercise-induced muscle damage and dysferlin is little reported.
OBJECTIVE: To observe the cell membrane permeability and expression levels of dysferlin and calpain3 in the rat gastrocnemius after acute eccentric exercise, so as to provide an theoretical reference for exploring the molecular mechanism of muscle regeneration and the exercise therapy of muscular diseases.
METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups including control, 24, 48 and 72 hours post exercise groups. The membrane permeability and expression levels of dysferlin and calpain3 were determined by immunohistochemistry, western blot assay and qRT-PCR.
RESULTS AND CONCLUSION: The activity of serum creatine kinase, and expression levels of calpain3 mRNA and dysferlin protein, as well as membrane permeability at 24 hours post exercise were significantly greater than those in the control group (P < 0.05 or 0.01). The expression level of dysferlin mRNA at 24 and 48 hours post exercise was significantly higher than that at 72 hours post exercise (P < 0.05). Therefore, the damage to the skeletal muscle cell membrane was the most severe at 24 hours after eccentric exercise. Due to Ca2+ influx, expression of calpain3 mRNA was activated, and then the damaged cell membrane was repaired by increasing the expression of dysferlin.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Tissue Engineering, Sports Medicine, Cell Membrane

CLC Number:

R318

Huang Qiao-ting1, 2, Xu Jie1, Lin Jia-shi3. Mechanism by which dysferlin promotes repair of exercise-induced skeletal muscie injury in rats[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(16): 2502-2507.

Figures/Tables 1

2.1 实验动物数量分析纳入大鼠32只，无死亡或感染，均进入结果分析。 2.2　离心运动后大鼠血清肌酸激酶活性的变化高强度间歇离心运动后，大鼠血清肌酸激酶活性逐渐升高，运动后24 h组与对照组相比，差异有显著性意义(P < 0.01)。随后血清肌酸激酶活性逐渐下降，至运动后72 h，略低于对照组，但组间差异无显著性意义。详细情况见图1。 2.3 Dysferlin蛋白免疫荧光染色结果对照组dysferlin荧光信号较弱，主要分布在骨骼肌细胞膜附近，且分布较为均匀。高强度间歇训练后，dysferlin荧光信号逐渐增强，虽然也主要分布在肌纤维膜周围，但在细胞内可以明显地观测到呈弥散性分布的荧光信号，以运动后24，72 h组尤为显著。见图2。 2.4 间歇性离心运动对大鼠腓肠肌dysferlin及calpain3 mRNA表达的影响与对照组相比，间歇性离心运动后dysferlin mRNA表达量持续增加，于运动后48 h达到峰值，随后逐渐下降至略低于对照组水平。运动后48 h组dysferlin mRNA表达量与运动后72 h相比较差异具有非常显著性意义(P < 0.01)；运动后24 h组dysferlin mRNA表达量与运动后72 h相比较差异有显著性意义(P < 0.05)。Calpain3 mRNA表达水平在运动后24 h达到峰值，与对照组相比差异有显著性意义(P < 0.01)，随后逐渐下降，见图3。 2.5 间歇性离心运动后大鼠腓肠肌dysferlin及calpain3蛋白表达变化采用蛋白免疫印迹法对各组大鼠腓肠肌dysferlin及calpain3蛋白表达量进行测定，经过图像分析软件处理后得出积分吸光度值，以对照组积分吸光度值平均数作为基数，其他组积分吸光度值除以基数作为蛋白变化结果，如图4所示。运动后24，48，72 h组dysferlin蛋白表达量均高于对照组，但仅有24 h组与对照组相比，具有显著性差异，而各组大鼠calpain3蛋白表达量均差异无显著性意义。"

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