中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (23): 3714-3720.doi: 10.12307/2023.487

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

有氧耐力运动调控大鼠心肌细胞自噬的机制

刘吉焕1，王  鹏1，袁顺灵1，简  晔1，黄少泽1，姚思思1，刘文锋1，2   

  1. 1湖南师范大学体适能与运动康复湖南省重点实验室，湖南省长沙市  410012；2湖南师范大学蛋白质化学与发育生物学教育部重点实验室，湖南省长沙市  410081
  • 收稿日期:2022-02-23 接受日期:2022-07-18 出版日期:2023-08-18 发布日期:2023-01-16
  • 通讯作者: 刘文锋，博士，教授，湖南师范大学体适能与运动康复湖南省重点实验室，湖南省长沙市 410012；湖南师范大学蛋白质化学与发育生物学教育部重点实验室，湖南省长沙市 410081
  • 作者简介:刘吉焕，男，1995年生，山西省乡宁县人，汉族，湖南师范大学在读硕士，主要从事运动与脑健康方面的研究。
  • 基金资助:
    国家自然科学基金项目(81702236)，项目负责人：刘文锋；湖南省教育厅重点项目(20A333)，项目负责人：刘文锋；湖南省自然科学基金项目(2018JJ3363)，项目负责人：刘文锋；湖南省高校青年骨干教师项目(湘教通[2020]43号)，项目负责人：刘文锋；湖南省研究生科研创新项目(湘教通[2021] 267号)，项目负责人：刘吉焕；湖南师范大学青年优秀人才培养计划(ET1507)，项目负责人：刘文锋

Mechanism by which aerobic endurance exercise regulates cardiomyocyte autophagy in rats

Liu Jihuan1, Wang Peng1, Yuan Shunling1, Jian Ye1, Huang Shaoze1, Yao Sisi1, Liu Wenfeng1, 2   

  1. 1Hunan Provincial Key Laboratory of Physical Fitness and Exercise Rehabilitation, Hunan Normal University, Changsha 410012, Hunan Province, China; 2Key Laboratory of Protein Chemistry and Developmental Biology, Ministry of Education, Hunan Normal University, Changsha 410081, Hunan Province, China
  • Received:2022-02-23 Accepted:2022-07-18 Online:2023-08-18 Published:2023-01-16
  • Contact: Liu Wenfeng, MD, Professor, Hunan Provincial Key Laboratory of Physical Fitness and Exercise Rehabilitation, Hunan Normal University, Changsha 410012, Hunan Province, China; Key Laboratory of Protein Chemistry and Developmental Biology, Ministry of Education, Hunan Normal University, Changsha 410081, Hunan Province, China
  • About author:Liu Jihuan, Master candidate, Hunan Provincial Key Laboratory of Physical Fitness and Exercise Rehabilitation, Hunan Normal University, Changsha 410012, Hunan Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81702236 (to LWF); Key Project of Hunan Provincial Department of Education, No. 20A333 (to LWF); Natural Science Foundation of Hunan Province, No. 2018JJ3363 (to LWF); Hunan Province College Young Backbone Teacher Project, No. [2020]43 (to LWF); Hunan Provincial Postgraduate Research and Innovation Project, No. [2021]267 (to LJH); Young Talents Training Program of Hunan Normal University, No. ET1507 (to LWF)

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摘要：

文题释义：

细胞自噬：是真核生物中进化保守的对细胞内物质进行周转的重要过程。该过程中一些损坏的蛋白或细胞器被双层膜结构的自噬小泡包裹后，送入溶酶体(动物)或液泡(酵母和植物)中进行降解并得以循环利用。
有氧运动：是指主要以有氧代谢提供运动中所需能量的运动方式。运动负荷与耗氧量呈线性关系。

背景：近年来，衰老相关的慢性病研究在世界范围内受到广泛关注，心脏随着年龄增长可能会出现过早衰老，运动作为一种安全健康的干预措施可以延缓其衰老。
目的：观察实施规律有氧运动对心肌细胞增龄性老化的影响并探讨其机制。
方法：纳入SD大鼠68只，3月龄(青年)20只，13月龄 (中年)和22月龄(老年)各24只，每个年龄组再随机均分为静息组及运动组。对3个运动组实施为期10周的递增负荷有氧跑台运动，其中早期递增负荷6周，晚期恒负荷4周；3个静息组不进行有氧运动干预。干预结束后采用实时荧光定量PCR和免疫组化法检测心肌Beclin-1、CaMKⅡα1、AMPKα1/pAMPKα1和PI3K/Akt/mTOR等通路相关蛋白与基因的表达。

结果与结论：①实施规律有氧运动后，与青、中、老3个年龄的静息组相比，3个年龄的运动组大鼠心肌细胞自噬Beclin-1表达水平均下调；同时3个年龄的运动组大鼠心肌CaMKⅡα1 mRNA表达水平也下调；②3个年龄静息组pAMPKα1和AMPKα1蛋白表达水平均呈现增龄性趋势；青年、中年和老年运动组的AMPKα1表达水平分别上调16.50%(Ρ < 0.05)、63.09%(Ρ < 0.01)和31.12%(Ρ < 0.01)；pAMPKα1表达水平也显著上调，其中青年运动组为 131.29%(Ρ < 0.01)，中年运动组上调40.55%(Ρ < 0.01)，老年运动组上调16.77%(Ρ < 0.05)；③整体来看，与3个年龄静息组相比，3个年龄运动组大鼠心肌PI3K表达水平均上调，青年、中年运动组AKT1 mRNA表达水平均下调；关于mTOR mRNA表达水平，青年运动组稍下调，中年运动组变化不大，而老年运动组显著上调；④提示有氧运动可以显著上调老年组大鼠PI3K的表达，促进mTOR的活性，下调CaMKⅡα1的表达，下调pAMPKα1/AMPKα1的比值，抑制Beclin-1的表达，调控心肌自噬水平，使其趋向于正常稳定状态，从而延缓心肌衰老。

https://orcid.org/0000-0002-9187-7187(刘吉焕)

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

关键词: 规律有氧运动, 心肌, 老化, 细胞自噬, AMPK/pAMPK信号通路

Abstract: BACKGROUND: In recent years, studies on chronic diseases related to aging have attracted wide attention worldwide. As the heart grows with age, premature aging may occur. Exercise is a safe and healthy intervention that can delay its aging.
OBJECTIVE: To observe the effect of regular aerobic exercise on the aging of cardiomyocytes and explore its mechanism.
METHODS: Healthy male SPF Sprague-Dawley rats aged 3 months (young, n=20), 13 months (middle age, n=24), and 22 months (old, n=24) were randomly divided into youth rest group, youth exercise group, middle age rest group, middle age exercise group, old age rest group, and old age exercise group. The three resting groups did not do aerobic exercise intervention, and the three exercise groups were given a 10-week incremental load aerobic treadmill exercise consisting of 6-week incremental load exercise and 4-week constant load exercise. RT-PCR and immunohistochemical detection were used to detect myocardial Beclin-1, CaMKIIα1, AMPKα1/pAMPKα1, and PI3K/Akt/mTOR pathway-related proteins and genes. 
RESULTS AND CONCLUSION: After regular aerobic exercises, compared with the three resting groups, the expression levels of Beclin-1 in cardiomyocytes were significantly down-regulated in the three exercise groups. The expression levels of myocardial CaMKIIα1 mRNA were also down-regulated in the three exercise groups. The protein expression levels of pAMPKα1 and AMPKα1 in the three resting groups showed an aging trend, and the expression levels of AMPKα1 in the young, middle age, and old age exercise groups were up-regulated by 16.50% (P < 0.05), 63.09% (P < 0.05), and 31.12% (P < 0.01), respectively. The expression levels of pAMPKα1 were also significantly up-regulated by 131.29% in the young exercise group (P < 0.01), 40.55% in the middle age exercise group (P < 0.01), and 16.77% in the old age exercise group (P < 0.05). Overall, the expression levels of PI3K in the myocardium were down-regulated in the three exercise groups compared with the three resting groups, while the expression levels of AKT1 mRNA were down-regulated in the young and middle age exercise groups. The expression level of mTOR mRNA was slightly down-regulated in the young exercise group, little changed in the middle age exercise group, and significantly up-regulated in the old age exercise group. To conclude, aerobic exercise can significantly up-regulate the expression of PI3K, promote the activity of mTOR, down-regulate the expression of CaMKIIα1, down-regulate the ratio of pAMPKα1/AMPKα1, decrease the expression of Beclin-1, regulate the level of myocardial autophagy, and make it tend to be normal and stable in aged rats, thereby delaying myocardial aging.

Key words: regular aerobic exercise, myocardium, aging, autophagy, AMPK/pAMPK signaling pathway

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