外源性重组膜联蛋白A6对小鼠离心运动后骨骼肌早期质膜损伤修复的影响

关键词：离心运动  重组人源膜联蛋白A6  骨骼肌损伤  质膜修复

Keywords:eccentric exercise  recombinant human Annexin A6  skeletal muscle injury  plasma membrane repair

摘要：

      摘要 目的：探究外源性重组人源膜联蛋白A6（recombinant human Annexin A6，rhANX A6）干预对离心运动损伤模型小鼠骨骼肌早期质膜损伤的影响及其代谢特点。 方法：将56只8周龄C57雄性小鼠随机分为安静对照组（C）、运动注射磷酸缓冲盐溶液（phosphate buffer saline，PBS）组（EP）和运动注射rhANX A6组（ER），其中运动组根据运动后不同取材时间点分为EP2、EP6、EP12和ER2、ER6、ER12，每组各8只。运动前2h根据小鼠体重对EP组和ER组分别腹腔注射1倍浓度PBS缓冲液和rhANX A6（0.8mg/kg）。运用ELISA检测各组小鼠血清肌酸激酶（creatine kinase，CK）、膜联蛋白A6（Annexin A6，ANX A6）及骨骼肌中ANX A6的含量，运用伊文思蓝染色（Evans blue dye，EBD）观察各组小鼠骨骼肌质膜的损伤情况。 结果：①EP组各时相、ER12h血清CK活力值显著高于C组（P<0.01）；同一时间点对比，EP组各时相均显著高于ER组（P<0.05）。②EP组及ER组各时相EBD阳性区域平均荧光强度均显著高于C组（P<0.01）；EP组各时相均显著高于ER组（P<0.01）。③ER组各时相及EP6h、EP12h血清ANX A6含量显著高于C组（P<0.01）；12h时相ER组显著高于EP组（P<0.05）。④ER组及EP组各时相骨骼肌ANX A6显著高于C组（P<0.05）；ER组各时相均显著高于EP组（P<0.05）。 结论：小鼠离心运动前2h注射外源性rhANX A6可显著改善运动后骨骼肌早期质膜损伤，推测rhANX A6直接参与了离心运动诱导的骨骼肌早期质膜损伤修复。

Abstract:

      Abstract Objective： To investigate the effect of exogenous recombinant human Annexin A6 （rhANX A6） intervention on early plasma membrane damage and its metabolic characteristics in skeletal muscle in mice with eccentric exercise injury models. Method： Fifty-six C57 male mice, aged 8 weeks, were randomly divided into the quiet control group （group C）， exercise group injected with phosphate-buffer saline (group EP） and exercise group injected with rhANX A6（group ER）. The exercise groups were further subdivided based on different post-exercise sampling times: EP2， EP6， EP12， and ER2， ER6， ER12，with 8 mice in each subgroup. Two hours before exercise, the group EP and group ER received intraperitoneal injections with 1*PBS buffer (1 × concentration) and rhANX A6 （0.8 mg/kg）, respectively. Elisa was used to detect the content of ANX A6 in serum creatine kinase （CK）， Annexin A6 （ANX A6） and skeletal muscle in each group of mice， and Evans Blue dye （EBD） was used to observe the damage of the skeletal muscle plasma membrane of each group of mice. Result： The serum CK activity in all EP subgroups and ER12h were significantly higher than those in the C group （P<0.01）. At the same time point， CK levels in all EP subgroups were significantly higher than in the ER group （P<0.05）. The average fluorescence intensity of EBD-positive regions in all EP and ER subgroup was significantly higher than that in group C （P<0.01）. All EP subgroups had significantly higher intensities than the ER group （P<0.01）. The serum ANX A6 levels in all ER subgroups and EP6h, EP12h were significantly higher than that in group C （P<0.01）. At 12h time point, ANX 46 levels in the ER group were significantly higher than in the EP group（P<0.05）. ANX A6 levels in skeletal muscle of all EP and ER subgroups were significantly higher than that in group C （P<0.05）； All ER subgroups had significantly higher than those in the EP group （P<0.05）. Conclusion： Exogenous rhANX A6 injection 2 hours before eccentric exercise in mice can significantly improve the early plasma membrane injury in skeletal muscle after exercise. It is speculated that rhANX A6 directly involved in the repair of early plasma membrane damage in skeletal muscle induced by eccentric exercise.