Cell apoptosis and expression of malondialdehyde in the skeletal muscle in a model rat undergoing long-term and high-intensity exercise

doi:10.3969/j.issn.2095-4344.0834

Abstract

Abstract:

BACKGROUND: Apoptosis in the skeletal muscle leads to a variety of skeletal muscle diseases, but there are ideal no therapies so far.
OBJECTIVE: To analyze the effects of different intensities of exercises on the morphology and apoptosis of the skeletal muscle in rats.
METHODS: Thirty healthy male 3-month-old Sprague-Dawley rats were randomly allocated into three groups (n=10 per group): control group (no swimming), aerobic exercise and high-intensity exercise groups. The rats in the exercise groups underwent swimming for consecutive 10 weeks. Then, the gastrocnemius muscle was isolated from all rats under anesthesia. The morphology and cell apoptosis were detected by hematoxylin-eosin staining and TUNEL test. At the same time, the levels of superoxide dismutase and malondialdehyde in skeletal muscle tissue were tested and the correlation of malondialdehyde with integrated optical density (IOD) was analyzed.
RESULTS AND CONCLUSION: The results of hematoxylin-eosin staining showed that the muscle fibers were in disorder arrangement with obscure structure in the high-intensity exercise group. Compared with the control group, the content of malondialdehyde was increased in the exercise groups, especially in the aerobic exercise group (P < 0.05). The level of superoxide dismutase in the high-intensity exercise group was significantly lower than that in the aerobic exercise group (P < 0.05). The results of TUNEL test showed that the IOD values in the high-intensity exercise group were significantly higher than those in the aerobic exercise and control groups (P < 0.05). There was a positive correlation of malondialdehyde with IOD. These results indicate that the long-term and high-intensity exercise can increase the content of malondialdehyde in the skeletal muscle, resulting in an aggravation of fatigue. Aerobic exercise can obtain the adaptation of the skeletal muscle, but damage the morphology and structure of the skeletal muscle because of excessive exercise load, thus accelarating the cell apoptosis. With the exercise intensity increasing, the balance between malondialdehyde production and clearance is broken, and the IOD value is increased.

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: Muscle, Skeletal, Apoptosis, Tissue Engineering

CLC Number:

R318

Ma Xue, Hu Ya-zhe. Cell apoptosis and expression of malondialdehyde in the skeletal muscle in a model rat undergoing long-term and high-intensity exercise[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(12): 1928-1933.

Figures/Tables 2

2.1 实验动物数量分析实验选用大鼠30只，分为3组，实验过程死亡5只，进入结果分析25只。 2.2 不同干预组对大鼠骨骼肌形态结构的影响安静对照组骨骼肌组织结构正常，肌纤维紧密的排列，形状规则；一般有氧运动组运动组肌纤维变粗，肌细胞间隙变大；长时间大强度运动组损伤处肌纤维锐性断裂成小片段，多数断面整齐，断间端连接紧密，损伤严重处断片排列紊乱结构模糊，即细胞溶解的结果。见图1。 2.3 不同干预组对大鼠骨骼肌超氧化物歧化酶、丙二醛的影响 ①丙二醛：一般有氧运动组与安静对照组相比，P < 0.05(t=2.742，P=0.014 5)；长时间大强度运动组与安静对照组相比，P < 0.01(t=7.266，P=0.000 1)；长时间大强度运动组与一般有氧运动组相比，P < 0.05 (t=2.887，P=0.012)；②超氧化物歧化酶：一般有氧运动组与安静对照组相比，P < 0.05(t=2.350，P =0.034 0)；长时间大强度运动组与一般有氧运动组相比，P < 0.01(t=3.574，P=0.003 1)。 2.4 不同强度有氧运动对大鼠骨骼肌细胞凋亡的影响 2.4.1 TUNEL检测显微镜下观察结果安静对照组可见少量阳性表达的细胞核；一般有氧运动组阳性表达细胞核数量相对多；长时间大强度运动组阳性表达的细胞核数量越多。见图2。 2.4.2 TUNEL染色显微图像分析积分吸光度结果一般有氧运动组与安静对照组相比，P < 0.05(t=2.617，P=0.018 7)；长时间大强度运动组与安静对照组相比，P < 0.01(t=6.905，P=0.000 1)；长时间大强度运动组与一般有氧运动组相比，P < 0.01 (t=4.574，P=0.004)。结果见表2。 2.5 丙二醛与积分吸光度值的相关性分析结果认为丙二醛与积分吸光度之间存显著相关，并且呈现正相关关系(r=0.702，P < 0.01)，即随着丙二醛浓度的增高，积分吸光度值增大。说明丙二醛对细胞凋亡有重要的作用。"

References

[1] Kerr JF, Wyllie AH, Currie AR.Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer.1972;26: 239-257.

[2] Schoser BG, Wehling S, Blottner D. Cell death and apoptosis-related proteins in muscle biopsies of sporadic amyotrophic lateral sclerosis and polyneuropathy. Muscle Nerve. 2001;24:1083-1089.

[3] 郑澜,潘珊珊,张胜年.骨骼肌细胞凋亡研究进展[J].中国运动医学杂志,2000,19(3):301-303.

[4] 谢昆,李兴权. 细胞凋亡的信号通路[J]. 山东农业大学学报:自然科学版,2015,46(04):514-518.

[5] 张颖, 梁美杨. 8周有氧运动对大鼠骨骼肌细胞凋亡的影响[J].哈尔滨体育学院学报, 2015,33(2):87-91.

[6] 张梦颖,吕志跃,吴忠道.细胞凋亡发生机制研究进展[J].热带医学杂志,2016,16(10):1346-1349+1352.

[7] 韩奇鹏,罗玲,揭红东,等. 细胞凋亡及其调控因子研究进展[J]. 饲料博览,2016,(4):9-13.

[8] 蔡保塔,余斌,赵江萍,等. 不同运动强度与骨骼肌细胞凋亡的时序性实验研究[J]. 南方医科大学学报,2006,26(7):1017-1019.

[9] 刘丽萍. 细胞凋亡与运动训练关系的研究进展[J]. 河北体育学院学报, 2000,14(1):75-80.

[10] 李莎,缪律,林缘西. 运动训练与细胞自噬、细胞凋亡的分子机制[J]. 南京体育学院学报:自然科学版,2016,(1):27-34.

[11] 金其贯. 慢性力竭性训练对大鼠骨骼肌细胞凋亡的影响[J].体育与科学,1999,20(5):23-28+56.

[12] 金其贯,冯美云.运动与细胞凋亡的研究进展[J]. 西安体育学院学报, 2001,21(3):30-34+40.

[13] Tezini GC, Silveira LC, Maida KD, et al. The effect of ovariectomy on cardiac autonomic control in rats submitted to aerobic physical training. Auton Neurosci. 2008;143(1):5-11.

[14] 胡亚哲,王姝玉,王和平,等.不同强度运动大鼠心肌细胞凋亡及一氧化氮合酶变化[J]. 中国运动医学杂志,2007,26(2): 222-225+ 254.

[15] 田振军,熊正英,郭进,等.大鼠游泳过度训练模型的建立及心室构型改建研究[J]. 陕西师范大学学报:自然科学版,1996,(3): 108-113.

[16] 王爱莲,史晓燕.统计学[M].西安:西安交通大学出版社, 2010: 212.

[17] 谢康玲,刘遂心,蔡颖,等. 长期中等强度运动对小鼠骨骼肌HIF-1α mRNA的表达及葡萄糖转运的影响[J].中国康复医学杂志, 2012,27(6):514-518.

[18] 刘军. Ebselen预处理对大强度耐力训练大鼠骨骼肌结构和功能的保护及可能机制研究[J].体育科学,2014,34(8):91-98.

[19] 陈英杰,郭庆芳.延迟性肌肉损伤与自由基代谢异常[J].中国运动医学杂志, 1993, 12(2): 65-69.

[20] 赵甜甜. 紫甘薯花青素对过度训练大鼠骨骼肌氧化损伤保护作用的研究[D].江苏师范大学,2013

[21] Gore M, Fiebig R, Hollander J. Endurance training alters antioxidant enzyme gene expression in rat skeletal muscle. Can J Physiol Pharmacol.1998;76(12):1139-1145.

[22] 聂园园,仇小强. 心肌细胞凋亡与心血管疾病关系的研究进展[J].实用医学杂志,2012,28(2):324-326.

[23] 袁海燕,胡亚哲.不同强度有氧运动对大鼠肝细胞的影响[J].中国组织工程研究,2015,19(51):8311-8316.

[24] Munsch D, Watanabe-Fukunaga R, Bourdon JC, et al.Human and mouse Fas (APO-1/CD95) death receptor genes each contain a p53-responsive element that is activated by p53 mutants unable to induce apoptosis. J Biol Chem.2000;275(6): 3867-3872.

[25] Volk T, Hensel M, Kox WJ. Transient Ca2+ changes in endothelial cells induced by low doses of reactive oxygen species: role of hydrogen peroxide. Mol Cell Biochem.1997; 171(1-2):11-21.

[26] 任小双,张兰,张燕宁,等. 昆虫细胞凋亡的线粒体通路研究进展[J]. 安徽农业科学,2017,45(3):167-170+189.

[27] 向军军,赖菁菁,胡跃强. 近3年线粒体介导细胞凋亡的研究进展[J]. 中西医结合心脑血管病杂志,2016,14(13):1497-1499.

[28] 李兴太,刘德文,张新,等. 线粒体通透性转换与细胞凋亡[J].中国公共卫生,2015,31(4):529-532.

[29] 王佺荃,张文丽,元小冬,等. 线粒体在细胞凋亡中的作用[J]. 中国医学创新,2015,12(06):143-146.

[30] Cain K, Bratton SB, Langlais C, et al. Apaf-1 oligomerizes into biologically active approximately 700-kDa and inactive approximately1.4-MDa apoptosome complexes. J Biol Chem. 2000;275(9):6067-6070.

[31] Attisano L, Wrana JL. Signal transduction by the TGF-beta superfamily. Science.2002;296(5573):1646-1647.

[32] Munsch D, Watanabefukunaga R, Bourdon J, et al. Human and Mouse Fas (APO-1/CD95) Death Receptor Genes Each Contain a p53-responsive Element That Is Activated by p53 Mutants Unable to Induce Apoptosis. J Biol Chem. 2000; 275(6):3867-3872.

[33] 王长青,李雷,郑师陵. 游泳训练后大鼠骨骼肌细胞自由基代谢、线粒体膜电位变化与细胞凋亡的关系[J]. 中国运动医学杂志, 2002,21(3): 256-260.

[34] 黄文聪,阮利民,王安利.不同负荷游泳训练对大鼠骨骼肌HSP70应答及其脂质过氧化的影响[J]. 体育科学,2008,28(3):63-66+ 78.

[35] 李烨,王逸安,张君,等. 丙二醛对体外培养下小鼠骨髓间充质干细胞凋亡的诱导作用[J]. 生命科学研究,2014,18(2):129-133.

[36] Munsch D, Watanabefukunaga R, Bourdon J, et al. Human and Mouse Fas (APO-1/CD95) Death Receptor Genes Each Contain a p53-responsive Element That Is Activated by p53 Mutants Unable to Induce Apoptosis. J Biol Chem. 2000; 275(6):3867-3872.

[37] 韩宁. 大鼠脑出血模型中神经细胞凋亡与自由基水平的相关性研究[D].第二军医大学,2007.