过度疲劳状态模型大鼠心脏功能及心肌细胞凋亡调控因子的变化
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摘要
背景:力竭运动是超出人或动物生理极限的运动,强烈的运动刺激会打破机体原有的稳态,从而引起一系列应激反应,过度应激会出现损伤和功能异常。目的:通过大鼠心肌细胞运动性疲劳模型分析导致运动性心脏猝死的相关因素。方法:130只SD大鼠购自成都达硕生物科技公司。随机选取7只大鼠作为空白组对照,其余大鼠为疲劳模型,对大鼠进行游泳训练,每36 h训练1次,连续训练致过度疲劳。建模成功后出现超量恢复的负叠加状态即为过度疲劳状态,并分别训练3×36 h,6×36 h,9×36 h后,随机处死各7只大鼠,分别记录为疲劳组A组,疲劳组B组和疲劳组C组;训练过程中或训练结束24h死亡的大鼠(排除因呛水死亡的大鼠)记为运动性猝死组。TUNEL法检测大鼠心肌细胞凋亡;免疫组织化学方法心肌组织Bax、Bcl-2表达;苏木精-伊红染色观察细胞形态变化。结果与结论:①在猝死组中,心肌纤维与其余组相比较为纤细,部分肌纤维出现断裂,血管极度扩张,心肌出现严重坏死伴随充血出血;②疲劳组及猝死组心肌细胞凋亡指数、心肌细胞凋亡数量都出现明显的增长(P <0.05);③当处于过度疲劳状态时,大鼠心脏组织中的促凋亡蛋白数量明显增加(P <0.05),抑制凋亡蛋白Bcl-2表达明显下降(P <0.01);④结果提示,连续训练而导致大鼠处于过度疲劳状态,会引发大鼠心肌细胞形态结构方面的变化,甚至导致损害,此时心肌组织中所含有的Bcl-2和Bax蛋白会出现异常表达情况。由于心肌细胞受到损害而导致心肌细胞凋亡,数量明显减少,进而影响整体心脏的结构以及功能,最终导致心源性运动性猝死。
BACKGROUND: Exhaustive exercise is a movement that exceeds the physiological limits of humans or animals. High-intensity motor exercise can disturb the homeostasis, causing a series of stress reactions, and excessive stress can lead to damage and dysfunction. OBJECTIVE: To analyze the related factors to sudden cardiac death by exercise-induced fatigue model of rat cardiomyocytes. METHODS: One hundred and thirty Sprague-Dawley rats were provided by Chengdu Dashuo Biotechnology Co., Ltd. Seven rats were randomly selected to as blank control group, and the remaining rats were used to establish the fatigue model(continuous swimming training, 36 hours per times). Then the negative superposition state of over-recovery was the state of over-fatigue. Seven rats were sacrificed and recorded as fatigue groups A, B and C after 3 x36 hours, 6 x36 hours and 9 x36 hours of training, respectively. The rats died during training or within 24 hours after training(excluding inhaled water) regarded as exercise-induced sudden death group. Apoptosis of cardiomyocytes was detected by TUNEL. Bax and Bcl-2 expression in myocardium was detected by immunohistochemistry. Cell morphology was observed by hematoxylin-eosin staining. RESULTS AND CONCLUSION:(1) In the sudden death group, the myocardial fibers were thinner than those in the other groups, some myocardial fibers were broken, the vessels were extremely dilated, and the myocardium was severely necrotic with hyperemia and hemorrhage.(2) Apoptotic index of cardiomyocytes and number of apoptotic cardiomyocytes in the fatigue and sudden death groups were significantly increased(P < 0.05).(3) The number of pro-apoptotic proteins in rat heart tissues were increased significantly in the state of excessive fatigue(P < 0.05), and the content of the apoptotic inhibiting protein Bcl-2 was decreased significantly(P < 0.01).(4) These results imply that continuous training can lead to excessive fatigue in rats, which will lead to changes in the morphological structure of rat cardiac cells and even lead to damage. Meanwhile, abnormal expression of Bcl-2 and Bax proteins in the cardiac tissue will occur. The damage of cardiomyocytes leads to apoptosis of cardiomyocytes, and the number of cardiomyocytes is significantly reduced, which affects the structure and function of the whole heart and eventually leads to sudden cardiac death.
BACKGROUND: Exhaustive exercise is a movement that exceeds the physiological limits of humans or animals. High-intensity motor exercise can disturb the homeostasis, causing a series of stress reactions, and excessive stress can lead to damage and dysfunction. OBJECTIVE: To analyze the related factors to sudden cardiac death by exercise-induced fatigue model of rat cardiomyocytes. METHODS: One hundred and thirty Sprague-Dawley rats were provided by Chengdu Dashuo Biotechnology Co., Ltd. Seven rats were randomly selected to as blank control group, and the remaining rats were used to establish the fatigue model(continuous swimming training, 36 hours per times). Then the negative superposition state of over-recovery was the state of over-fatigue. Seven rats were sacrificed and recorded as fatigue groups A, B and C after 3 x36 hours, 6 x36 hours and 9 x36 hours of training, respectively. The rats died during training or within 24 hours after training(excluding inhaled water) regarded as exercise-induced sudden death group. Apoptosis of cardiomyocytes was detected by TUNEL. Bax and Bcl-2 expression in myocardium was detected by immunohistochemistry. Cell morphology was observed by hematoxylin-eosin staining. RESULTS AND CONCLUSION:(1) In the sudden death group, the myocardial fibers were thinner than those in the other groups, some myocardial fibers were broken, the vessels were extremely dilated, and the myocardium was severely necrotic with hyperemia and hemorrhage.(2) Apoptotic index of cardiomyocytes and number of apoptotic cardiomyocytes in the fatigue and sudden death groups were significantly increased(P < 0.05).(3) The number of pro-apoptotic proteins in rat heart tissues were increased significantly in the state of excessive fatigue(P < 0.05), and the content of the apoptotic inhibiting protein Bcl-2 was decreased significantly(P < 0.01).(4) These results imply that continuous training can lead to excessive fatigue in rats, which will lead to changes in the morphological structure of rat cardiac cells and even lead to damage. Meanwhile, abnormal expression of Bcl-2 and Bax proteins in the cardiac tissue will occur. The damage of cardiomyocytes leads to apoptosis of cardiomyocytes, and the number of cardiomyocytes is significantly reduced, which affects the structure and function of the whole heart and eventually leads to sudden cardiac death.
引文
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[8]万思源,蔡传元,张艳青,等.左甲状腺素和维生素E对甲状腺功能减退症大鼠心肌损伤的影响[J].安徽医科大学学报, 2013,48(1):21-24.
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[10]刘峰.姜黄素对力竭运动大鼠心肌抗氧化、糖代谢及细胞凋亡的影响[J].扬州大学学报(农业与生命科学版),2017,38(2):40-46.
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[12]张龙飞,崔玉娟,平政,等.红景天苷对力竭大鼠心肌线粒体呼吸功能的影响[J].解放军医药杂志,2014,26(11):1-5.
[13]徐鹏,康亭,曹雪滨,等.力竭运动后不同时相大鼠心电图、心功能变化及Nrf2的作用[J].中国应用生理学杂志, 2016,32(2):146-151.
[14]公雪,李晓燕.力竭运动对心肌组织结构及功能影响的研究进展[J].心血管病学进展,2016,37(4):435-440.
[15]赵小辉.红景天苷对急性力竭运动大鼠的心脏保护作用及机制研究[J].中国妇幼健康研究,2016,27(2):261.
[16]王永良,刘忠民,刘眉眉,等.不同负荷游泳运动对老龄大鼠心肌细胞形态及Bax、Bcl-2蛋白表达的影响[J].中国老年学杂志, 2017,4(37):1857-1859.
[17]Chatard JC,Mujika I,Goiriena JJ,etal.Screening young athletes for prevention of sudden cardiac death:Practical recommendations for sports physicians.Scand J MedSci Sports.2016;26(4):362-374.
[18]费萍燕,彭生,费民忠,等.miR7在心肌细胞缺氧复氧模型中的表达及对心肌细胞凋亡的影响[J].使用预防医学,2018,25(2):195-198.
[19]孙晓娟,力竭运动损伤模型大鼠运动预适应后心脏STAT3和Caspase-3的表达[J].中国组织工程研究, 2015,19(40):6450-6454.
[20]田阁,徐昕,李国平.C/EBPβ在预防心源性运动猝死中的潜在作用[J].中国运动医学杂志,2018,37(3):263-266.
[2]莫轶,韦军.我国运动心脏的诊断现状及进展[J].中国临床新医学,2018,11(02):208-212.
[3]殷劲,岳建兴,周进等.糖酵解供能运动后“超量恢复区间”的研究[J].成都体育学院学报,2007,33(4):88-91.
[4]Carbone A,D'andrea A,Riegler L,etal. Cardiac damage in athlete's heart:When the"supernormal"heart fails.World J Cardiol.2017;9(6):470-480.
[5]钱钰,殷维瑶,李华.过度疲劳状态下运动性猝死模型大鼠甲状腺及心脏功能变化[J].中国组织工程研究, 2016,20(49):7356-7363.
[6]钱钰,李华,殷维瑶.过度疲劳状态下运动性猝死大鼠甲状腺功能的变化[J].中国体育科技,2016,6(52):92-98.
[7]阳仁均,殷维瑶,李华.力竭运动性猝死模型大鼠运动皮质Bax、Bcl-2及脑源性神经营养因子的表达变化[J].中国组织工程研究,2016,20(49),7377-7383.
[8]万思源,蔡传元,张艳青,等.左甲状腺素和维生素E对甲状腺功能减退症大鼠心肌损伤的影响[J].安徽医科大学学报, 2013,48(1):21-24.
[9]Oláh A, Németh BT, Mátyás C, et alCardiac effects of acute exhaustive exercise in a rat model. Int J Cardiol. 2015;182:258.
[10]刘峰.姜黄素对力竭运动大鼠心肌抗氧化、糖代谢及细胞凋亡的影响[J].扬州大学学报(农业与生命科学版),2017,38(2):40-46.
[11]翟帅,陈佩林.运动心脏重塑过程中miRNA-350/JNK信号通路的动态变化[J].体育科学,2014,34(5):9-14.
[12]张龙飞,崔玉娟,平政,等.红景天苷对力竭大鼠心肌线粒体呼吸功能的影响[J].解放军医药杂志,2014,26(11):1-5.
[13]徐鹏,康亭,曹雪滨,等.力竭运动后不同时相大鼠心电图、心功能变化及Nrf2的作用[J].中国应用生理学杂志, 2016,32(2):146-151.
[14]公雪,李晓燕.力竭运动对心肌组织结构及功能影响的研究进展[J].心血管病学进展,2016,37(4):435-440.
[15]赵小辉.红景天苷对急性力竭运动大鼠的心脏保护作用及机制研究[J].中国妇幼健康研究,2016,27(2):261.
[16]王永良,刘忠民,刘眉眉,等.不同负荷游泳运动对老龄大鼠心肌细胞形态及Bax、Bcl-2蛋白表达的影响[J].中国老年学杂志, 2017,4(37):1857-1859.
[17]Chatard JC,Mujika I,Goiriena JJ,etal.Screening young athletes for prevention of sudden cardiac death:Practical recommendations for sports physicians.Scand J MedSci Sports.2016;26(4):362-374.
[18]费萍燕,彭生,费民忠,等.miR7在心肌细胞缺氧复氧模型中的表达及对心肌细胞凋亡的影响[J].使用预防医学,2018,25(2):195-198.
[19]孙晓娟,力竭运动损伤模型大鼠运动预适应后心脏STAT3和Caspase-3的表达[J].中国组织工程研究, 2015,19(40):6450-6454.
[20]田阁,徐昕,李国平.C/EBPβ在预防心源性运动猝死中的潜在作用[J].中国运动医学杂志,2018,37(3):263-266.