耐力运动训练对大鼠骨骼肌线粒体能量代谢的影响
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摘要
目的：通过检测大鼠骨骼肌腺苷酸含量,线粒体呼吸链氧化酶(COX、NOX)活性和抗氧化能力(SOD、GSH活性和MDA含量),阐明不同强度的耐力运动训练对骨骼肌腺苷酸含量,线粒体呼吸链氧化酶的活性以及骨骼肌抗氧化能力的影响规律,从线粒体呼吸链氧化酶活性的运动适应性变化和骨骼肌抗氧化能力变化两个角度探索不同强度的耐力运动训练后骨骼肌出现能量代谢适应性变化的可能机制。方法：将40只SD雌性大鼠参照Bedford报道大鼠最大摄氧量的标准,随机分为安静组(C)、小强度组(L)、中等强度(M)和大强度组H),每组10只,即：安静组(C)：常规饲养不运动；大、中、小强度组训练两天,休息一天,再训练三天,休息一天。共持续7周；实验结束时将大鼠按组分批处理取大鼠内侧腓肠肌和比目鱼肌。HPLC法测定骨骼肌ATP、ADP和AMP含量；分光光度法测定NADH、CCO氧化酶活性和抗氧化酶SOD、GSH活性和MDA含量。
结果：
1.腺苷酸方面：腓肠肌的ATP、AMP值均高于比目鱼肌的ATP、AMP值,比目鱼肌的ADP值均高于腓肠肌。小强度训练不但使腓肠肌ATP、AMP含量和比目鱼肌的ATP、ADP和能荷含量显著提高(P<0.05),也使腓肠肌的能荷非常显著提高(P<0.01)。中强度训练不但使腓肠肌的ATP、AMP含量、能荷和比目鱼肌的ATP、ADP含量非常显著提高(P<0.01),也显著提高了比目鱼肌的能荷(P<0.05)。大强度训练后,腓肠肌的ATP较安静组显著下降(P<0.05),能荷降低非常显著(P<0.01)。
2.呼吸链氧化酶方面：小强度训练使比目鱼肌的NOX、COX活性提高非常显著(P<0.01)。中强度训练不但使腓肠肌的NOX、COX活性和比目鱼肌的NOX活性显著提高(P<0.05),也非常显著提高了比目鱼肌的COX活性(P<0.01)。大强度训练后,腓肠肌的NOX、COX活性提高非常显著(P<0.01)。
3.抗氧化能力方面：小强度训练使比目鱼肌的GSH活性显著提高(P<0.05), SOD非常显著提高(P<0.01),腓肠肌的MDA含量显著提高(P<0.05)。中强度训练不但使腓肠肌的SOD、GSH活性、MDA含量和比目鱼肌的SOD活性显著提高(P<0.05)。大强度训练后,腓肠肌的SOD、GSH活性提高非常显著(P<0.01),而比目鱼肌的MDA含量提高显著(P<0.05)。
4.比目鱼肌能荷与COX、NOX活性存在非常显著或显著的正相关性(r=0.752),((r=0.844),比目鱼肌ATP与COX活性存在非常显著的正相关性(r=0.716)。慢肌的能荷升高受NOX、COX活性调节,且COX活性更敏感一些,而ATP的升高则主要受COX活性的调节。
5.比目鱼肌能荷与GSH、SOD活性存在显著或非常显著的高度正相关性(r=0.720)(r=0.925),比目鱼肌ATP与SOD活性存在非常显著的正相关性(r=0.773)。慢肌的能荷升高受SOD、GSH活性调节,且SOD活性更敏感一些,而ATP的升高则主要受SOD活性的调节。
6.比目鱼肌的NOX与SOD、GSH活性具有显著性正相关(r=0.793)(r=0.726),比目鱼肌的COX与SOD、GSH活性具有显著或非常显著性正相关(r=0.843)(r=0.669),腓肠肌COX与GSH具有显著的中度正相关(r=0.586)。
结论：
1.小、中强度除对骨骼肌的ATP,能荷有较大提高外,对慢肌的ADP敏感性较高,而对快肌的AMP敏感性较高。
2.小强度训练显著提高慢肌的呼吸链氧化酶活性和抗氧化能力；大强度显著提高快肌的呼吸链氧化酶活性和抗氧化能力。
3.耐力训练可以通过提高SOD和GSH活性,保证细胞膜的完整,提高呼吸链氧化酶的活性,使骨骼肌出现能量代谢适应性提高,尤其是慢肌的线粒体能量代谢。
Purpose:Through detecting the content of adenosine acid, the activity of oxidase of mitochondria respiratory chain (COX, NOX)and the anti-oxidation capacity (the activity of SOD, GSH and the content of MDA) in skeletal muscle of rats, we expound the influence rule of the content of adenosine acid,the activity of oxidase of mitochondria respiratory chain, and the oxidation resistance ability in the skeletal muscle during different intensity endurance movement training. From the standpoint of the change of running suitability of the activity of oxidase of mitochondria respiratory chain and the anti-oxidation capacity, we investigate possible mechanism of adaptive changes in energy metabolism of the skeleton muscle; after different intensity endurance trainings.
Methods:In this study, refering to the standard of maximal oxygen intake Bedford reported, 40 SD female big mouse were used and divided randomly into four groups:(1)Control-group (C), (2)low-intensity group (L), (3)medium -intensity (M), (4)high-intensity group (H).Each one has 10.Control-group rats were fed normally. Other -group rats set training two day, take a rest for a day, train again for three days and take a rest for a day. It sustains totally 7 cycles. Soleus (SOL) and medial grastrocnemius (MG) of rats in groups and batches were handled after experiment. Determination of ATP, ADP, AMP, in Skeletal Muscle by HPLC; The activities of cytochrome oxidase(CCO), NADH oxidase and the activities of antioxidase SOD, GSH-PX, and MDA were measured by Spectrophotometry.
Results:
1. Adenosine acid:the levels of ATP、AMP of rat gastrocnemius muscle are higher than that of the soleus muscle. The levels of ADP of rat soleus muscle are higher than that of the gastrocnemius muscle. The levels of ATP、AMP of rat gastrocnemius muscle and the levels of ATP、ADP、EC of rat soleus muscle in LG were significantly higher than that in CG (P<0.05). The levels of EC of rat gastrocnemius muscle in LG were significantly higher than that in CG very much (P<0.01). The levels of ATP、AMP、EC of rat gastrocnemius muscle and the levels of ATP、ADP of rat soleus muscle in MG were significantly higher than that in CG very much (P<0.01). The levels of EC of rat soleus muscle in MG were significantly higher than that in CG (P<0.05). The levels of ATP,EC of rat gastrocnemius muscle in HG were significantly decreased than that in CG (P<0.05～0.01).
2. Oxidase of mitochondria respiratory chain:the levels of NOX、COX of rat soleus muscle in LG were significantly higher than that in CG (P<0.05). The levels of NOX、COX of rat gastrocnemius muscle and the levels of NOX of rat soleus muscle in MG were significantly higher than that in CG very much (P<0.05). The levels of COX of rat soleus muscle in MG were significantly higher than that in CG very much (P<0.01). The levels of NOX、COX of rat gastrocnemius muscle in HG were significantly higher than that in CG very much (P<0.01).
3. The anti-oxidation capacity:The levels of MDA of rat gastrocnemius muscle and the levels of GSH、SOD of rat soleus muscle in LG were significantly higher than that in CG (P<0.05-0.01). The levels of SOD、GSH、MDA of rat gastrocnemius muscle and the levels of SOD of rat soleus muscle in MG were significantly higher than that in CG (P<0.05). The levels of SOD、GSHof rat gastrocnemius muscle in HG were significantly higher than that in CG very much (P<0.01). The levels of MDA rat soleus muscle in HG were significantly higher than that in CG (P<0.05).
4. The contents of EC of rat soleus muscle were very positive significant correlated with COX (P< 0.01) and NOX (P< 0.05). The contents of ATP of rat soleus muscle were very positive significant correlated with COX (P< 0.01).The activity of EC of slow muscle can be regulated by NOX、COX, and the COX activeness is more sensitive,but The activity of ATP of slow muscle can be regulated by COX.
5. The activity of NOX of rat soleus muscle was very positive significant correlated with SOD and GSH (P< 0.05). The activity of COX of rat soleus muscle was very positive significant correlated with SOD (P< 0.05) and GSH (P< 0.01). The activity of COX of rat gastrocnemius muscle was very positive significant correlated with GSH (P< 0.05).
Conclusions:
1. Except to the activity of ATP, EC of skeletal muscle was sensitive to LG and CG training,the activity of ADP was sensitive to slow muscle in LG and CG, but the activity of AMP was sensitive to fast muscle in LG and CG.
2. The activity of oxidase of mitochondria respiratory chain and the anti-oxidation capacity in slow muscle of rats in LG were obviously improved. The activity of oxidase of mitochondria respiratory chain and the anti-oxidation capacity in fast muscle of rats in HG were obviously improved.
3. After endurance sports training, the activity of SOD and GSH were obviously improved, so that the integrity of cell membrane was assured. The changes of CCO activity is closely related to mitochondrial energy metabolism enhancement, particularly mitochondrial energy metabolism in slow muscle.
结果：
1.腺苷酸方面：腓肠肌的ATP、AMP值均高于比目鱼肌的ATP、AMP值,比目鱼肌的ADP值均高于腓肠肌。小强度训练不但使腓肠肌ATP、AMP含量和比目鱼肌的ATP、ADP和能荷含量显著提高(P<0.05),也使腓肠肌的能荷非常显著提高(P<0.01)。中强度训练不但使腓肠肌的ATP、AMP含量、能荷和比目鱼肌的ATP、ADP含量非常显著提高(P<0.01),也显著提高了比目鱼肌的能荷(P<0.05)。大强度训练后,腓肠肌的ATP较安静组显著下降(P<0.05),能荷降低非常显著(P<0.01)。
2.呼吸链氧化酶方面：小强度训练使比目鱼肌的NOX、COX活性提高非常显著(P<0.01)。中强度训练不但使腓肠肌的NOX、COX活性和比目鱼肌的NOX活性显著提高(P<0.05),也非常显著提高了比目鱼肌的COX活性(P<0.01)。大强度训练后,腓肠肌的NOX、COX活性提高非常显著(P<0.01)。
3.抗氧化能力方面：小强度训练使比目鱼肌的GSH活性显著提高(P<0.05), SOD非常显著提高(P<0.01),腓肠肌的MDA含量显著提高(P<0.05)。中强度训练不但使腓肠肌的SOD、GSH活性、MDA含量和比目鱼肌的SOD活性显著提高(P<0.05)。大强度训练后,腓肠肌的SOD、GSH活性提高非常显著(P<0.01),而比目鱼肌的MDA含量提高显著(P<0.05)。
4.比目鱼肌能荷与COX、NOX活性存在非常显著或显著的正相关性(r=0.752),((r=0.844),比目鱼肌ATP与COX活性存在非常显著的正相关性(r=0.716)。慢肌的能荷升高受NOX、COX活性调节,且COX活性更敏感一些,而ATP的升高则主要受COX活性的调节。
5.比目鱼肌能荷与GSH、SOD活性存在显著或非常显著的高度正相关性(r=0.720)(r=0.925),比目鱼肌ATP与SOD活性存在非常显著的正相关性(r=0.773)。慢肌的能荷升高受SOD、GSH活性调节,且SOD活性更敏感一些,而ATP的升高则主要受SOD活性的调节。
6.比目鱼肌的NOX与SOD、GSH活性具有显著性正相关(r=0.793)(r=0.726),比目鱼肌的COX与SOD、GSH活性具有显著或非常显著性正相关(r=0.843)(r=0.669),腓肠肌COX与GSH具有显著的中度正相关(r=0.586)。
结论：
1.小、中强度除对骨骼肌的ATP,能荷有较大提高外,对慢肌的ADP敏感性较高,而对快肌的AMP敏感性较高。
2.小强度训练显著提高慢肌的呼吸链氧化酶活性和抗氧化能力；大强度显著提高快肌的呼吸链氧化酶活性和抗氧化能力。
3.耐力训练可以通过提高SOD和GSH活性,保证细胞膜的完整,提高呼吸链氧化酶的活性,使骨骼肌出现能量代谢适应性提高,尤其是慢肌的线粒体能量代谢。
Purpose:Through detecting the content of adenosine acid, the activity of oxidase of mitochondria respiratory chain (COX, NOX)and the anti-oxidation capacity (the activity of SOD, GSH and the content of MDA) in skeletal muscle of rats, we expound the influence rule of the content of adenosine acid,the activity of oxidase of mitochondria respiratory chain, and the oxidation resistance ability in the skeletal muscle during different intensity endurance movement training. From the standpoint of the change of running suitability of the activity of oxidase of mitochondria respiratory chain and the anti-oxidation capacity, we investigate possible mechanism of adaptive changes in energy metabolism of the skeleton muscle; after different intensity endurance trainings.
Methods:In this study, refering to the standard of maximal oxygen intake Bedford reported, 40 SD female big mouse were used and divided randomly into four groups:(1)Control-group (C), (2)low-intensity group (L), (3)medium -intensity (M), (4)high-intensity group (H).Each one has 10.Control-group rats were fed normally. Other -group rats set training two day, take a rest for a day, train again for three days and take a rest for a day. It sustains totally 7 cycles. Soleus (SOL) and medial grastrocnemius (MG) of rats in groups and batches were handled after experiment. Determination of ATP, ADP, AMP, in Skeletal Muscle by HPLC; The activities of cytochrome oxidase(CCO), NADH oxidase and the activities of antioxidase SOD, GSH-PX, and MDA were measured by Spectrophotometry.
Results:
1. Adenosine acid:the levels of ATP、AMP of rat gastrocnemius muscle are higher than that of the soleus muscle. The levels of ADP of rat soleus muscle are higher than that of the gastrocnemius muscle. The levels of ATP、AMP of rat gastrocnemius muscle and the levels of ATP、ADP、EC of rat soleus muscle in LG were significantly higher than that in CG (P<0.05). The levels of EC of rat gastrocnemius muscle in LG were significantly higher than that in CG very much (P<0.01). The levels of ATP、AMP、EC of rat gastrocnemius muscle and the levels of ATP、ADP of rat soleus muscle in MG were significantly higher than that in CG very much (P<0.01). The levels of EC of rat soleus muscle in MG were significantly higher than that in CG (P<0.05). The levels of ATP,EC of rat gastrocnemius muscle in HG were significantly decreased than that in CG (P<0.05～0.01).
2. Oxidase of mitochondria respiratory chain:the levels of NOX、COX of rat soleus muscle in LG were significantly higher than that in CG (P<0.05). The levels of NOX、COX of rat gastrocnemius muscle and the levels of NOX of rat soleus muscle in MG were significantly higher than that in CG very much (P<0.05). The levels of COX of rat soleus muscle in MG were significantly higher than that in CG very much (P<0.01). The levels of NOX、COX of rat gastrocnemius muscle in HG were significantly higher than that in CG very much (P<0.01).
3. The anti-oxidation capacity:The levels of MDA of rat gastrocnemius muscle and the levels of GSH、SOD of rat soleus muscle in LG were significantly higher than that in CG (P<0.05-0.01). The levels of SOD、GSH、MDA of rat gastrocnemius muscle and the levels of SOD of rat soleus muscle in MG were significantly higher than that in CG (P<0.05). The levels of SOD、GSHof rat gastrocnemius muscle in HG were significantly higher than that in CG very much (P<0.01). The levels of MDA rat soleus muscle in HG were significantly higher than that in CG (P<0.05).
4. The contents of EC of rat soleus muscle were very positive significant correlated with COX (P< 0.01) and NOX (P< 0.05). The contents of ATP of rat soleus muscle were very positive significant correlated with COX (P< 0.01).The activity of EC of slow muscle can be regulated by NOX、COX, and the COX activeness is more sensitive,but The activity of ATP of slow muscle can be regulated by COX.
5. The activity of NOX of rat soleus muscle was very positive significant correlated with SOD and GSH (P< 0.05). The activity of COX of rat soleus muscle was very positive significant correlated with SOD (P< 0.05) and GSH (P< 0.01). The activity of COX of rat gastrocnemius muscle was very positive significant correlated with GSH (P< 0.05).
Conclusions:
1. Except to the activity of ATP, EC of skeletal muscle was sensitive to LG and CG training,the activity of ADP was sensitive to slow muscle in LG and CG, but the activity of AMP was sensitive to fast muscle in LG and CG.
2. The activity of oxidase of mitochondria respiratory chain and the anti-oxidation capacity in slow muscle of rats in LG were obviously improved. The activity of oxidase of mitochondria respiratory chain and the anti-oxidation capacity in fast muscle of rats in HG were obviously improved.
3. After endurance sports training, the activity of SOD and GSH were obviously improved, so that the integrity of cell membrane was assured. The changes of CCO activity is closely related to mitochondrial energy metabolism enhancement, particularly mitochondrial energy metabolism in slow muscle.
引文
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