人工模拟不同低氧训练对大鼠心肌线粒体呼吸链酶复合物活性及自由基代谢的影响
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摘要
传统的高原训练利用高原缺氧和运动双重刺激,使运动员产生激烈的应激反应,以调动体内的机能潜力,提高运动能力。随着高原训练研究的发展,高原训练的基础理论研究不断完善;训练方法不断改进,科学化程度不断提高,从而逐步建立了一些新的训练手段及模拟训练方法。本实验利用低压氧舱设备模拟不同的低氧训练方法,建立动物训练模型,运用生理学和生物化学理论和方法,对大鼠心肌线粒体呼吸链酶活性及自由基代谢相关指标进行统计分析,探讨不同低氧训练模式对线粒体呼吸链酶活性及自由基代谢相关指标的影响,希望能为广大教练员和运动员在选择低氧训练模式时提供相关理论依据。
实验方法:健康雄性大鼠40只,体重120±20g,所有大鼠在适应性饲养16d后,按要求随机分成低住低练组(LoLo 8只),高住高练组(HiHi 8只),高住低训组(HiLo 8只),低住高练组(LoHi 8只)和高住高练低训组(HiHiLo 8只)。根据慢性运动性疲劳动物训练模型,实验动物进行递增负荷的跑台训练,每天训练1次,每周6天,一共5周。坡度为零。在训练过程中,低氧训练第一周速度为20m/min、第二、三、四周速度为25m/min,五周速度为30m/min,常氧训练第一周速度为25m/min,第二、三、四周速度为30m/min,第五周速度为35m/min,训练时间都从30min递增到60min。训练5周后,休息两天,然后所有实验大鼠一次性跑台运动至力竭后即刻断头处死,分别测定各组大鼠心脏重量指数、心肌线粒体呼吸链酶复合物Ⅰ-Ⅳ活性和心肌线粒体SOD活性及MDA含量。
实验结果:
①至实验结束时,各低氧组与对照组相比,各组大鼠体重都呈明显增长趋势,常氧训练组、低住高练组以及高住高练组大鼠体重增长相对较快,略高于高住高练低训组和高住低训组,但各组间大鼠体重变化不具有统计学意义(P>0.05)。
②与常氧训练组比较,5周实验结束时各低氧训练组大鼠心脏重量指数都无显著性差异(P>0.05);HiHiLo组心指数较对照组略有提高但不具有统计学意义(P>0.05)。
③与常氧训练组比较, HiHiLo和HiHi组力竭时间较对照组略有提高但不具有统计学意义(P>0.05);LoHi和HiLo组力竭时间比对照组稍短(P>0.05)。
④与常氧训练组比较,HiHi组大鼠跑台至力竭后心肌线粒体SOD活性虽有升高但不具有统计学意义(P>0.05),而MDA含量则显著性降低(P<0.05);HiLo组线粒体SOD活性有一定的升高,但没有显著性差异(P>0.05),而MDA含量则明显低于LoLo组(P<0.05);LoHi组线粒体SOD活性、MDA含量均低于对照组,但均无显著性差异(P﹥0.05); HiHiLo组SOD活性有非常显著性升高(P<0.01);而MDA含量则明显低于LoLo组(P<0.05)。
⑤与LoLo组比较,在力竭运动后即刻,HiHi组大鼠心肌线粒体呼吸链酶CⅠ、CⅡ活性略有升高,但无统计学意义(P﹥0.05),CⅢ活性显著性降低(P<0.05); CⅣ活性显著性升高(P﹤0.05);HiLo组CⅠ、CⅣ活性比LoLo组低,但无差异性显著(P﹥0.05),CⅡ、CⅢ活性略有升高,但无统计学意义(P﹥0.05); LoHi组CⅠ、CⅡ活性都有升高,但无统计学意义(P﹥0.05); CⅢ、CⅣ活性均比LoLo组低,且CⅢ活性具有显著性差异(P﹤0.05); HiHiLo组CⅠ活性有极显著性提高(P﹤0.001),CⅡ活性也显著性提高(P﹤0. 01),CⅢ活性显著性降低(P﹤0.05), CⅣ活性差异达到了极显著性水平(P﹤0.001)。
结论:
①不同低氧训练对大鼠体重增长没有显著性影响。
②不同低氧训练对大鼠心脏重量指数没有显著影响;但HiHiLo模式对心指数的提高具有积极性意义。
③不同低氧训练对大鼠一次性跑台运动至力竭的时间没有显著影响;但HiHi、HiHiLo模式对大鼠耐力性提高具有积极意义。
④不同低氧训练在一定程度上可以提高大鼠心脏线粒体SOD活性、降低MDA含量; HiHiLo、HiHi、HiLo模式对提高机体抗氧化能力具有积极意义。
⑤低氧结合运动训练可以在一定程度上对大鼠心肌线粒体呼吸链酶复合物活性产生良性影响,加速线粒体氧化磷酸化过程,为线粒体合成ATP提供条件,但不同低氧训练模式对心肌线粒体呼吸链酶复合物活性的影响因部位不同而影响程度不同。
The traditional plateau training takes use of double stimulations of training and hypoxia environment, athletes will be able to endure severe stimulations to mobilize body potential, to arouse a series of anti一hypoxia physiological and biochemical reactions. With the develop- ment of altitude training researches, the researches of basic theories under altitude training are perfectly being built up, training methods have been more improved, scientific trainings have been more enhanced.Thus some new training means and stimulant training methods have been established. This experiment ,which make use of hypoxia cabin equipment simulation different hypoxia training method ,building-up animal training method ,applied theory and method of athletic physiology and sports biochemistry statistical to analyze the activity of respiratory complexesⅠ,Ⅱ,Ⅲ,Ⅳand free radicals metabolism in mitochondria.Explore the effect of activities of respiratory chain in mitochondria and free radicals metabolism of mice’s heart when different hypoxia training methods put on them .And hope for providing the certain theoretical reference For coaches and athletes when they want to choose hypoxia training methods . Experimental methods:
40male healthy mice(weight 120±20g) were randomly divided into 5 groups after 16days adaptive feeding. They are Living Low-Training Low(LoLo number8),Living High Training High(HiHi number8),Living High Training Low (HiLo number8),Living Low Training High(LoHi number8),Living High Training High exercise High(HiHiLo number8). According to chronic fatigue animal movement of the training model, experimental animals all carry on gradually intensive pedestal training once a day, sum up five weeks. Zero slope. In the training process, the first week of training hypoxia speed of 20 m / min, the second ,third and fourth week is 25 m / min, the fifth week is 30 m / min, the first week of normal training is 25 m / min, the second ,third and fourth week is 30 m / min, the fifth week is 35 m / min, training time increased from 30 min to 60 min. After Training five weeks ,have a rest for two days, and then all the rats were killed by decollation after exhaustive , the rats were measured heart weight/bodyweight, Superoxide dismutase(SOD)and malondialdehyde(MDA) in mitochondria,and activity of respiratory complexesⅠ,Ⅱ,ⅢandⅣwere measured in mitochondria isolated from heart.
Result of the experiment:
①After 5 weeks’feeding and training , all rats showed significant growth trend of body weight;The weight of mice from LoLo,LiHo and HiHi increased faster than HiHiLo and HiLo relatively . but the wight among groups did not change in significantly (P> 0.05).
②Compared with LoLo, The heart wight / body wight of hypoxia group showed no significant difference(P > 0.05) ;The HiHiLo’heart wight / body wight increased relatively, But no remarkable difference (P > 0.05).
③Compared with LoLo,The time of mice running to exhaustion of HiHiLo and HiHi showed a relative increase scope, but no obvious difference (P > 0.05); The time of mice running to exhaustion of LoHi and HiLo was a little shorter than LoLo’s(P > 0.05).
④Compared with the control group(LoLo),The activity of SOD in mitochondria of mice’s heart from HiHi showed a relative increase, but no significant difference after running to exhaustion(P > 0.05),and the amount of MDA decreased remarkably (P<0.05);The activity of SOD from HiLo represented a certain increase, but there is no significant difference between them (P> 0.05),The amount of MDA showed an obvious decrease(P <0.05); The activity of SOD and the amount of MDA of LoHi all showed a little lower dgree than loLo’s, but no distinct difference after exhaustive running (P> 0.05);The activity of SOD from HiHiLo showed an obvious increase(P <0.001);and the amount of MDA decreased significa- ntly(P<0.05).
⑤Compared with LoLo, The activity of CⅠ, CⅡin mitochondrial respiratory chain of mice’s heart from HiHi showed an increasing trend after exhaustive running,but there was no remarkable difference(P> 0.05), The activity of CⅢwas decreased significantly(P <0.05), and the activity of CⅣrevealed a significant rise (P <0.05); Both the activities of CⅠand CⅣfrom HiLo were lower than LoLo’s(P> 0.05); and the activities of CⅢand CⅡrepresent an increasing trend ,but no obvious difference(P> 0.05); The activities of CⅠ、CⅡof LoHi revealed a relative ascending, but no marked difference (P> 0.05),the activities of CⅢand CⅣwere lower than LoLo’,and CⅢpresented decreasing significantly(P <0.05);The activities of CⅠfrom HiHiLo showed an obvious increase (P <0.001),so did as the activity of CⅡ(P﹤0. 01),And CⅢdecreased significantly(P <0.05), CⅣrised remarkably after running to exhaustion (P <0.001) .
Conclusion:
①There was no remarkable effect of different hypoxia training methods on the mice’s weight.
②Although no remarkable effect of different hypoxia training methods on the mice’s heart weight to body weight, there was a positive sense in proving heart weight to body weight of HiHiLo.
③There was no effect of different hypoxia training methods on prolonging the time of mice running to exhaustion, but HiHiLo’s resistance was better than others.
④There was different increasing in activity of SOD and declining in the amount of MDA from hypoxia training methods .And the manifestation of the hypoxic group is not the same as different method; HiHiLo、HiHi and LoHi showed a better result than others.
⑤There are some positive impact on the mitochondrial respiratory chain of mice’s heart from hypoxic training and this could accelerate the process of mitochondrial oxidative phosphorylation and help to synthesize ATP, However, effect on parts of mitochondrial respiratory chain to be different because of different training method.
实验方法:健康雄性大鼠40只,体重120±20g,所有大鼠在适应性饲养16d后,按要求随机分成低住低练组(LoLo 8只),高住高练组(HiHi 8只),高住低训组(HiLo 8只),低住高练组(LoHi 8只)和高住高练低训组(HiHiLo 8只)。根据慢性运动性疲劳动物训练模型,实验动物进行递增负荷的跑台训练,每天训练1次,每周6天,一共5周。坡度为零。在训练过程中,低氧训练第一周速度为20m/min、第二、三、四周速度为25m/min,五周速度为30m/min,常氧训练第一周速度为25m/min,第二、三、四周速度为30m/min,第五周速度为35m/min,训练时间都从30min递增到60min。训练5周后,休息两天,然后所有实验大鼠一次性跑台运动至力竭后即刻断头处死,分别测定各组大鼠心脏重量指数、心肌线粒体呼吸链酶复合物Ⅰ-Ⅳ活性和心肌线粒体SOD活性及MDA含量。
实验结果:
①至实验结束时,各低氧组与对照组相比,各组大鼠体重都呈明显增长趋势,常氧训练组、低住高练组以及高住高练组大鼠体重增长相对较快,略高于高住高练低训组和高住低训组,但各组间大鼠体重变化不具有统计学意义(P>0.05)。
②与常氧训练组比较,5周实验结束时各低氧训练组大鼠心脏重量指数都无显著性差异(P>0.05);HiHiLo组心指数较对照组略有提高但不具有统计学意义(P>0.05)。
③与常氧训练组比较, HiHiLo和HiHi组力竭时间较对照组略有提高但不具有统计学意义(P>0.05);LoHi和HiLo组力竭时间比对照组稍短(P>0.05)。
④与常氧训练组比较,HiHi组大鼠跑台至力竭后心肌线粒体SOD活性虽有升高但不具有统计学意义(P>0.05),而MDA含量则显著性降低(P<0.05);HiLo组线粒体SOD活性有一定的升高,但没有显著性差异(P>0.05),而MDA含量则明显低于LoLo组(P<0.05);LoHi组线粒体SOD活性、MDA含量均低于对照组,但均无显著性差异(P﹥0.05); HiHiLo组SOD活性有非常显著性升高(P<0.01);而MDA含量则明显低于LoLo组(P<0.05)。
⑤与LoLo组比较,在力竭运动后即刻,HiHi组大鼠心肌线粒体呼吸链酶CⅠ、CⅡ活性略有升高,但无统计学意义(P﹥0.05),CⅢ活性显著性降低(P<0.05); CⅣ活性显著性升高(P﹤0.05);HiLo组CⅠ、CⅣ活性比LoLo组低,但无差异性显著(P﹥0.05),CⅡ、CⅢ活性略有升高,但无统计学意义(P﹥0.05); LoHi组CⅠ、CⅡ活性都有升高,但无统计学意义(P﹥0.05); CⅢ、CⅣ活性均比LoLo组低,且CⅢ活性具有显著性差异(P﹤0.05); HiHiLo组CⅠ活性有极显著性提高(P﹤0.001),CⅡ活性也显著性提高(P﹤0. 01),CⅢ活性显著性降低(P﹤0.05), CⅣ活性差异达到了极显著性水平(P﹤0.001)。
结论:
①不同低氧训练对大鼠体重增长没有显著性影响。
②不同低氧训练对大鼠心脏重量指数没有显著影响;但HiHiLo模式对心指数的提高具有积极性意义。
③不同低氧训练对大鼠一次性跑台运动至力竭的时间没有显著影响;但HiHi、HiHiLo模式对大鼠耐力性提高具有积极意义。
④不同低氧训练在一定程度上可以提高大鼠心脏线粒体SOD活性、降低MDA含量; HiHiLo、HiHi、HiLo模式对提高机体抗氧化能力具有积极意义。
⑤低氧结合运动训练可以在一定程度上对大鼠心肌线粒体呼吸链酶复合物活性产生良性影响,加速线粒体氧化磷酸化过程,为线粒体合成ATP提供条件,但不同低氧训练模式对心肌线粒体呼吸链酶复合物活性的影响因部位不同而影响程度不同。
The traditional plateau training takes use of double stimulations of training and hypoxia environment, athletes will be able to endure severe stimulations to mobilize body potential, to arouse a series of anti一hypoxia physiological and biochemical reactions. With the develop- ment of altitude training researches, the researches of basic theories under altitude training are perfectly being built up, training methods have been more improved, scientific trainings have been more enhanced.Thus some new training means and stimulant training methods have been established. This experiment ,which make use of hypoxia cabin equipment simulation different hypoxia training method ,building-up animal training method ,applied theory and method of athletic physiology and sports biochemistry statistical to analyze the activity of respiratory complexesⅠ,Ⅱ,Ⅲ,Ⅳand free radicals metabolism in mitochondria.Explore the effect of activities of respiratory chain in mitochondria and free radicals metabolism of mice’s heart when different hypoxia training methods put on them .And hope for providing the certain theoretical reference For coaches and athletes when they want to choose hypoxia training methods . Experimental methods:
40male healthy mice(weight 120±20g) were randomly divided into 5 groups after 16days adaptive feeding. They are Living Low-Training Low(LoLo number8),Living High Training High(HiHi number8),Living High Training Low (HiLo number8),Living Low Training High(LoHi number8),Living High Training High exercise High(HiHiLo number8). According to chronic fatigue animal movement of the training model, experimental animals all carry on gradually intensive pedestal training once a day, sum up five weeks. Zero slope. In the training process, the first week of training hypoxia speed of 20 m / min, the second ,third and fourth week is 25 m / min, the fifth week is 30 m / min, the first week of normal training is 25 m / min, the second ,third and fourth week is 30 m / min, the fifth week is 35 m / min, training time increased from 30 min to 60 min. After Training five weeks ,have a rest for two days, and then all the rats were killed by decollation after exhaustive , the rats were measured heart weight/bodyweight, Superoxide dismutase(SOD)and malondialdehyde(MDA) in mitochondria,and activity of respiratory complexesⅠ,Ⅱ,ⅢandⅣwere measured in mitochondria isolated from heart.
Result of the experiment:
①After 5 weeks’feeding and training , all rats showed significant growth trend of body weight;The weight of mice from LoLo,LiHo and HiHi increased faster than HiHiLo and HiLo relatively . but the wight among groups did not change in significantly (P> 0.05).
②Compared with LoLo, The heart wight / body wight of hypoxia group showed no significant difference(P > 0.05) ;The HiHiLo’heart wight / body wight increased relatively, But no remarkable difference (P > 0.05).
③Compared with LoLo,The time of mice running to exhaustion of HiHiLo and HiHi showed a relative increase scope, but no obvious difference (P > 0.05); The time of mice running to exhaustion of LoHi and HiLo was a little shorter than LoLo’s(P > 0.05).
④Compared with the control group(LoLo),The activity of SOD in mitochondria of mice’s heart from HiHi showed a relative increase, but no significant difference after running to exhaustion(P > 0.05),and the amount of MDA decreased remarkably (P<0.05);The activity of SOD from HiLo represented a certain increase, but there is no significant difference between them (P> 0.05),The amount of MDA showed an obvious decrease(P <0.05); The activity of SOD and the amount of MDA of LoHi all showed a little lower dgree than loLo’s, but no distinct difference after exhaustive running (P> 0.05);The activity of SOD from HiHiLo showed an obvious increase(P <0.001);and the amount of MDA decreased significa- ntly(P<0.05).
⑤Compared with LoLo, The activity of CⅠ, CⅡin mitochondrial respiratory chain of mice’s heart from HiHi showed an increasing trend after exhaustive running,but there was no remarkable difference(P> 0.05), The activity of CⅢwas decreased significantly(P <0.05), and the activity of CⅣrevealed a significant rise (P <0.05); Both the activities of CⅠand CⅣfrom HiLo were lower than LoLo’s(P> 0.05); and the activities of CⅢand CⅡrepresent an increasing trend ,but no obvious difference(P> 0.05); The activities of CⅠ、CⅡof LoHi revealed a relative ascending, but no marked difference (P> 0.05),the activities of CⅢand CⅣwere lower than LoLo’,and CⅢpresented decreasing significantly(P <0.05);The activities of CⅠfrom HiHiLo showed an obvious increase (P <0.001),so did as the activity of CⅡ(P﹤0. 01),And CⅢdecreased significantly(P <0.05), CⅣrised remarkably after running to exhaustion (P <0.001) .
Conclusion:
①There was no remarkable effect of different hypoxia training methods on the mice’s weight.
②Although no remarkable effect of different hypoxia training methods on the mice’s heart weight to body weight, there was a positive sense in proving heart weight to body weight of HiHiLo.
③There was no effect of different hypoxia training methods on prolonging the time of mice running to exhaustion, but HiHiLo’s resistance was better than others.
④There was different increasing in activity of SOD and declining in the amount of MDA from hypoxia training methods .And the manifestation of the hypoxic group is not the same as different method; HiHiLo、HiHi and LoHi showed a better result than others.
⑤There are some positive impact on the mitochondrial respiratory chain of mice’s heart from hypoxic training and this could accelerate the process of mitochondrial oxidative phosphorylation and help to synthesize ATP, However, effect on parts of mitochondrial respiratory chain to be different because of different training method.
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