外源性重组膜联蛋白A6在离心运动后小鼠骨骼肌早期质膜损伤修复中的作用  

The role of recombinant human annexin A6 in the repair of early plasma membrane damage in mouse skeletal muscle after eccentric exercise

作　　者：韩艳丽 许寿生[1,2] 周越 吴迎[1,2] Han Yanli;Xu Shousheng;Zhou Yue;Wu Ying(School of Sport Science,Beijing Sport University,Beijing 100084,China;Key Laboratory of Sports Stress Adaptation,State General Administration of Sports,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]国家体育总局运动应激适应重点实验室,北京100084

出　　处：《中国运动医学杂志》2024年第10期791-802,共12页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金项目(31771312);中央高校基本科研业务费专项资金资助项目(2020054)。

摘　　要：目的:探究外源性重组人源膜联蛋白A6(rh ANX A6)干预在离心运动损伤模型小鼠骨骼肌早期质膜修复中的作用。方法:56只8周龄C57雄性小鼠随机分为安静对照组(C组,n=8)、运动注射PBS组(EP组,n=24)和运动注射rhANX A6组(ER组,n=24),其中EP、ER组分别于运动后2 h、6 h、12 h三个时间点取材,记为EP2、EP6、EP12和ER2、ER6、ER12,每组各8只。运动前2 h根据小鼠体重对EP、ER组分别给予腹腔注射1íPBS缓冲液和rhANX A6(0.8 mg/kg)。EP组和ER组进行一次下坡跑离心运动,跑台坡度15°,初始速度5 m/min,以1 m/min的速度递增至17 m/min,运动60 min。运动结束后,于相应时间点取材,采用Elisa检测各组血清肌酸激酶(CK)水平;免疫荧光观察xin肌动蛋白结合重复内含物(Xirp)染色阳性面积,膜联蛋白A6(ANX A6)与膜联蛋白A1(ANX A1)、膜联蛋白A2(ANX A2)的共定位变化及ANX A6介导的质膜修复孔径大小。结果:1)与C组相比,EP组各时刻、ER12血清CK水平显著升高(P<0.01);同一时刻对比,EP组各时刻均显著高于ER组(P<0.05)。2)与C组相比,EP组及ER组各时刻Xirp染色阳性面积百分比均显著升高(P<0.05);EP组2h、12 h显著高于ER组(P<0.01)。3)与C组相比,EP组及ER组各时刻ANX A6与A1的共定位系数均显著上升(P<0.01);EP组6 h、12 h显著高于ER组(P<0.05)。4)与C组相比,EP组及ER组各时刻ANX A6与A2的共定位系数显著升高(P<0.01);EP组6h、12 h显著高于ER组(P<0.01)。5)与C组相比,EP组各时刻及ER组6 h、12 h的ANX A6与A1共定位荧光点显著升高(P<0.05);EP组6 h、12 h显著高于ER组(P<0.01)。6)与C组相比,EP组及ER组6 h、12 h的ANX A6与A2共定位荧光点显著升高(P<0.01);EP组6 h、12 h显著高于ER组(P<0.01)。7)离心运动损伤后6 h,ANX A1、A2主要参与ANX A6介导的10μm左右的“大创口”质膜修复。结论:运动前2 h注射外源性rhANX A6可显著促进离心运动后早期骨骼肌质膜损伤修复,对微米级“补丁”的形成具有积极作�Objective To explore the role of exogenous recombinant human annexin A6(rhANX A6)intervention in early plasma membrane repair of skeletal muscles in mice with eccentric motion injury.Methods Fifty-six 8-week-old C57 male mice were randomly divided into a resting control group(C,n=8),an exercise+PBS injection group(EP,n=24)and an exercise+rhANX A6 injection group(ER,n=24).Two hours before the exercise,both EP and ER groups were injected 1×PBS buffer and rhANX A6(0.8 mg/kg)intraperitoneally,respectively.Both EP and ER groups underwent a 60-min downhill centrifugal running on the running platform of 15°slope,with an initial speed of 5 m/min,increased by 1 m/min up to 17 m/min.At 2 h,6 h and 12 h after exercise,8 mice were executed to collect samples.The serum creatine kinase(CK)levels were detected using enzyme-linked immunosorbent assay,while the positive area of Xin actin-binding repeat-containing protein(XIRP),the co localization of annexin A6(ANX A6),annexin A1(ANX A1)and annexin A2(ANX A2),as well as the pore size of plasma membrane repair mediated by ANX A6 were observed using immunofluorescence.Results Compared with group C,a significant increase was observed in the average serum CK level at all sampling points in EP group and 12 h after exercise in ER group(P<0.01),with that of EP group significantly higher than ER group at all sampling points(P<0.05).Moreover,the percentages of Xirp staining positive area in EP group and ER group were significantly higher group C at all time points(P<0.05),with that of EP group 2 h and 12 h after exercise significantly higher than ER group(P<0.01).Meanwhile,the co-localization coefficients of ANX A6 and A1 increased significantly at all sampling points in both EP and ER groups(P<0.01),with those of EP group 6 h and 12 h after exercise significantly higher than ER group(P<0.01).What’s more,compared with group C,the co-localization fluorescence points of ANX A6 with A1 or A2 of EP group at all times and of ER group 6 h and 12 h after exercise increased significantly,with

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 

分 类 号：R873[医药卫生—运动医学]