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无氧运动诱导的斑马鱼代谢及相关酶的适应性变化
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摘要
本研究以斑马鱼为研究对象,测定了斑马鱼10 min无氧运动后8h内的肌肉糖原和乳酸的代谢变化;研究了4周无氧运动训练对斑马鱼行为、形态、生长、肌肉生化组分、运动后肌糖原和乳酸含量、肌肉代谢酶活性的影响。旨在探索斑马鱼的无氧运动代谢特征和对无氧运动训练的适应性变化,为进一步研究鱼类适应无氧运动训练的分子机制提供基础数据。
     在经过10 min无氧驱赶运动后,斑马鱼肌糖原含量下降了76.90%,但未观察到力竭现象,在随后的8h逐渐恢复,达到到静止水平时的66.27%。运动后肌乳酸急剧上升达到16.43±1.39μmol/g wet wt,是静止水平的2.62倍。在随后的8h恢复中乳酸含量逐渐下降,4h和8h时肌乳酸含量已经同静止水平无显著差异。乳酸清除速率(LCR)计算得1.16μmol/h·g,乳酸产生系数(LPC)为1.50。
     斑马鱼4周无氧运动训练后对照组和训练组死亡率分别为8.0%和11.3%。训练期间,训练组个体较对照组日常活跃程度降低,而更喜集群游动。
     4周训练后对照组斑马鱼的体长显著增加,而训练组虽然体长略增加,但与训练前比较未有显著差异。对照组体重4周后显著增加,增幅达22.47%,而训练组体重略有下降,和训练前没有显著性差异。对照组的肥满度显著增加,训练组却显著下降。
     4周无氧运动训练促使训练组斑马鱼较对照组肌糖原含量显著增加54.72%,而肌肉内总蛋白含量和水分含量对照组和训练组之间无显著差异。
     对照组和训练组斑马鱼在10 min无氧运动后,对照组肌乳酸含量显著高于训练组,而肌糖原含量均显著下降,对照组和训练组肌糖原含量降低绝对值分别为8.51μmol/g wet wt和10.33μmol/g wet wt,下降百分比分别为61.77%和48.48%。
     4周训练后斑马鱼肌乳酸脱氢酶(LDH)活性显著增高,柠檬酸合成酶(CS)活性显著降低,而己糖激酶(HK)、磷酸果糖激酶(PFK)、丙酮酸激酶(PK)、细胞色素C氧化酶(COX)活性未被作用。
     以上结果表:10 min无氧运动不足以促使斑马鱼力竭。斑马鱼对乳酸的耐受性较强,属于乳酸释放型鱼类,其运动后乳酸清除时间短,但清除速率一般。4周无氧运动训练显著降低了斑马鱼日常的活跃程度,促使其群聚性程度增加。训练组个体生长被显著抑制,形态更加“苗条”而利于运动。训练促使斑马鱼肌糖原含量显著增加,加强了运动持久能力,乳酸耐受能力增强,同时加强了无氧代谢能力。斑马鱼个体无氧运动能力整体提升。
To explore anaerobic exercise metabolic characteristics and adaption changes to anaerobic exercise training of zebrafish (Danio rerio) and supply basic data for future molecular mechanism research of adaption to anaerobic exercise training of fish, zebrafish was used to measure the muscle glycogen and lactate concentration changes in 8 h after 10 min anaerobic exercise. And also the influence of 4 weeks anaerobic exercise training on behavior, morphology, growth, biochemical components of muscle, muscle glycogen and lactate concentration following exercise and muscle metabolic enzyme activity were investigated.
     Muscle glycogen concentration of zebrafish reduced 76.90%, but no exhaustive behavior was observed following 10 min anaerobic chasing-exercise. Muscle glycogen concentration recovered regularly in 8 h, reached 66.27% of pre-exercise. Muscle lactate concentration increased sharply following exercise and reached 16.43±1.39μmol/g wet wt, it was 2.62 fold of pre-exercise. Muscle lactate concentration reduced regularly in 8 h and there was no significant difference at 4h and 8h compared to pre-exercise. The lactate clearance rate (LCR) is 1.16μmol/h·g and the lactate produce coefficient (LPC) is 1.50.
     The death rate of control and trained-group were 8.0%,11.3% respectively following 4 weeks anaerobic exercise training. The daily level of activity was less low in trained group than control, the trained zebrafish usually liked swimming together more than control.
     The training caused no significant increase in length and weight of trained group, but control zebrafish increased both the length and weight significantly. The condition factor of control increased significantly, but the trained group's condition factor was reduced significantly following training.
     4 weeks training induced significant increase in muscle glycogen of zebrafish, but training were no effects on protein and moisture of muscle.
     Muscle lactate concentration of control was significantly higher than trained group following 10 min anaerobic exercise and both groups of muscle glycogen were reduced, the decrease values of control and trained group were 8.51μmol/g wet wt and 10.33μmol/g wet wt respectively.
     4 weeks training significantly increased the activity of lactate dehydrogenase (LDH) in muscle, but decreased the activity of citrate synthase (CS). There were no effects on activity of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), cytochrome c oxidase (COX) in muscle.
     Results above indicate:10 min anaerobic exercise isn't able to cause exhaustion of zebrafish. Zebrafish is able to bear some degree concentration of lactate and belongs to“the lactate releaser”. Though the recovery time of post-exercise of zebrafish is short, the clearance rate is normal. The daily level of activity was declined by 4 weeks anaerobic training and they swim together more. The growth is inhibited significantly and zebrafish change slimmer adapted to locomotion. Training causes the increase of glycogen in muscle and enhances the exercise endurance and the ability of bearing lactate. And also the anaerobic metabolism is enhanced. The ability of anaerobic exercise of zebrafish is enhanced in general.
引文
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