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中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (3): 220-226.doi: 10.12047/j.cjap.6243.2022.041

• 研究论文 • 上一篇    下一篇

时钟基因BMAL1在运动性骨骼肌损伤恢复中的作用*

傅泽铤, 夏雨, 丁海丽   

  1. 成都体育学院, 四川 成都 610041
  • 收稿日期:2021-12-24 修回日期:2022-04-27 出版日期:2022-05-28 发布日期:2022-09-05
  • 通讯作者: Tel: 028-85051087; E-mail: dinghaili@.cdsu.edu.cn
  • 基金资助:
    *国家重点研发计划(2018YFF0300604);国家自然科学基金资助项目(81904318);运动医学四川省重点实验室暨国家体育总局重点实验室2021-A021

The role of clock gene BMAL1 in exercise-induced skeletal muscle injury recovery

FU Ze-ting, XIA Yu, DING Hai-li   

  1. Chengdu Sport University, Chengdu 610041, China
  • Received:2021-12-24 Revised:2022-04-27 Online:2022-05-28 Published:2022-09-05

摘要: 目的: 探讨时钟基因BMAL1在运动性骨骼肌损伤恢复中的作用。方法: 208只8周龄SD大鼠随机分为对照组(C组,n=104)和运动组(E组,n=104)。E组于跑台进行90 min下坡跑,运动后于0 h, 6 h,12 h,18 h, 24 h, 30 h, 36 h, 42 h, 48 h, 54 h, 60 h, 66 h, 72 h各取C组、E组8只大鼠腓肠肌,通过实时荧光定量PCR实验检测骨骼肌核心时钟基因BMAL1表达量,应用余弦分析软件circacompare (R package)获取拟合余弦曲线参数,分析其节律性振荡的变化趋势;透射电镜观察骨骼肌肌纤维超微结构变化;免疫印迹法(Western blot)检测骨骼肌BMAL1、DESMIN表达;免疫荧光观测BMAL1与DESMIN的定位及含量变化。结果: C组BMAL1 mRNA在72 h内呈现3个完整近日节律周期;E组BMAL1 mRNA在 0 h~24 h近日节律消失。与C组比较,E组在运动后0 h、6 h、12 h、18 h,BMAL1 mRNA含量显著升高(P<0.05),在运动后0 h、12 hBMAL1蛋白表达显著升高(P<0.05)、24 h至72 h恢复至无明显差异(P>0.05);在运动后0 h、12 h,DESMIN蛋白表达降低(P<0.05),24 h开始逐渐升高,48 h显著升高(P<0.01),至72 h恢复至无明显差异(P>0.05)。E组BMAL1与DESMIN在运动后0 h、12 h、24 h发生共定位,0 h~24 h共定位呈现先降低后升高的趋势,24 h的荧光强度达到最高值。结论: 运动后时钟基因BMAL1可能与调控骨架蛋白DESMIN的协同作用增强,从而与促进肌纤维结构恢复有关。

关键词: 大脑/肌肉芳香经受体核转位因子样蛋白1, 时钟基因, 运动性骨骼肌损伤, 结蛋白, 骨架蛋白, 大鼠

Abstract: Objective: To investigate the role of clock gene BMAL1 in exercise-induced skeletal muscle injury recovery. Methods: Two hundred and eight 8-week-old SD rats were randomly divided into the control group (Group C, n=104) and the exercise group (Group E, n=104). Group E performed a 90-minute downhill run on the treadmill. After exercise, the gastrocnemius muscle of 8 rats in Group C and Group E were collected at 0 h, 6 h, 12 h, 18 h, 24 h, 30 h, 36 h, 42 h, 48 h, 54 h, 60 h, 66 h and 72 h. The expression of skeletal muscle core clock gene, BMAL1 was measured by real-time fluorescence quantitative PCR. The parameters of fitting cosine curve were obtained by cosine analysis software circacompare (R package), and the change trend of rhythmic oscillation was analyzed. The ultrastructure of skeletal muscle fibers was observed by transmission electron microscope. The expressions of skeletal muscle BMAL1 and DESMIN were detected by Western blot; Immunofluorescence was used to observe the localization and contents of BMAL1 and DESMIN. Results: In Group C, three complete circadian rhythm cycles of mRNA BMAL1 were observed within 72 hours; in Group E, the circadian rhythm of BMAL1 mRNA disappeared at 0 h~24 h. Compared with Group C, the expression level of BMAL1 mRNA was significantly increased at 0 h, 6 h, 12 h, and 18 h after exercise in Group E (P<0.05), and the expression of BMAL1 protein was significantly increased at 0 h and 12 h after exercise(P<0.05), and recovered to the level of that in Group C from 24 h to 72 h(P>0.05). The expression of DESMIN protein was decreased at 0 h and 12 h after exercise(P<0.05), gradually increased at 24 h, increased significantly at 48 h(P<0.01), and recovered to the control level at 72 h (P>0.05). In Group E, BMAL1 and DESMIN were co-localized at 0 h, 12 h, and 24 h after exercise; the colocalization at 0 h~24 h showed a trend of first decreasing and then increasing, and the fluorescence intensity at 24 h reached the highest value. Conclusion: The post-exercise clock gene BMAL1 may be involved in the enhanced synergy of regulating the cytoskeletal protein DESMIN, it is thus related to the promotion of muscle fiber structure recovery.

Key words: brain and muscle ARNT- like1, clock gene, exercise-induced skeletal muscle injury, DESMIN, skeleton protein, rat

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