模拟不同高原训练方法对大鼠心肌超微结构及生理生化指标的影响
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摘要
本研究通过应用人工低压氧舱模拟各种不同高原训练方法,建立动物训练模型,应用形态学和生理生化检测方法,对不同训练模式下大鼠心肌及血液相关指标的测试及研究分析,对比研究各训练方式对心肌形态学和生化指标的影响,并从运动生物学和运动训练学的角度对这些方法的应用意义及存在的问题进行分析,以便帮助科研人员和教练员更全面地认识这些问题,同时在训练实践中更有效地利用这些方法。
按实验要求将大鼠随机分为低住安静组(LoC)、低住低练组(LoLo)、低住高练组(LoHi)、高住安静组(HiC)、高住低练组(HiLo)、高住高练组(HiHi),高住高练低训组(HiHiLo),共7组,每组10只。以A.X.Bigard跑台训练方法进行递增、大强度的跑台训练方式建立模拟低氧训练模型。各组大鼠在给予相应的模拟高原训练后(共4周),每组随机抽取一只断头处死,取心尖组织,制成50nm厚度切片,采用JEM2-1230型透射电镜技术从细胞核、线粒体、肌原纤维、心肌闰盘和毛细血管等五个方面观察大鼠心肌细胞超微结构在低氧训练过程中的变化。其余各组大鼠在模拟高原训练完成后,一次性在跑台跑至力竭,记录力竭时间,后断头处死,快速剥离心脏,取血液样品分离血清,将心肌组织和血清液氮冷冻,后置于冰箱(-40℃)保存。用试剂盒检测心肌组织和血清中的SOD、LDH、LA、CK、SDH、ATP酶的活性和含量。
实验结果表明:
1)对心肌线粒体的影响,其中LoC、LoLo、HiLo组的线粒体数目丰富、个数较多,结构完整无损伤;而LoHi、HiC、HiHi、HiHiLo组线粒体形态结构有不同程度的损伤,表现为体积大小不均,形态各异,有肿胀,嵴聚集,数目减少等。
2)对肌原纤维和肌细胞核的影响,LoC、LoLo、LoHi、HiLo组肌原纤维和肌细胞核结构完整,肌丝排列规则整齐,肌节清晰可见,肌膜完整;HiC、HiHi、HiHiLo组肌原纤维有变化,排列不整齐,长短不均一,部分局域肌丝缺失。
3)对心肌微血管和闰盘的影响,不同高原训练方式中HiHiLo组微血管有部分结构损伤,表现为血管内皮细胞核染色质聚集,肿胀膨大,内皮细胞有破坏。LoHi组心肌闰盘有变化,表现为桥粒减少、间隙增大、连接不紧密等。
4)对血清LA和LDH的影响,不同的低氧训练方法中对大鼠无氧代谢能力有不同的改善作用,其中以HiLo、LoHi和HiHi组值低于其他各训练组,提示其对乳酸有较好的耐受和清除能力。
5)对心肌ATP酶的影响,Na~+-K~+ATP酶以HiHiLo、HiLo组;Ca~(2+)、Mg~(2+)-ATPase以HiLo、HiHiLo、LoHi组酶活性优于其它各组。提示,主要的模拟高原训练方法HiLo、HiHiLo和LoHi组对心肌的ATP酶活性都有比较显著的改善作用。
6)心肌LDH值以HiHiLo组较高,心肌CK以HiLo、LoHi、HiHiLo组活性高于其他各组。提示,不同的低氧训练方法中对心肌无氧代谢改善作用比较显著的训练方法是HiLo、HiHiLo和LoHi组。
7)HiLo、LoHi法对心肌SOD活性的提高较其它各组显著。心肌有氧代谢酶SDH活性提高较显著的组是HiLo、LoHi组,提示模拟高原训练方法HiLo、LoHi训练法对运动大鼠心肌抗氧化能力和有氧代谢能力有较好的改善作用。。
结论:
1)经过4wk模拟不同高原训练,对心肌超微结构有不同的影响,HiLo组LoLo组心肌超微结构有适应性变化,而HiHiLo、HiHi组心肌结构在线粒体、微血管、肌纤维、心肌闰盘等方面表现有不同程度的损伤。提示,模拟高原训练方法对心肌超微结构的影响因训练方法的异同而产生不同的效果。
2)不同高原训练方式对心肌生理生化指标有不同影响,但整体来看,高原训练可以有效的改善机体机能,提高运动能力,不同的训练方法中尤以高住低练法较为显著,可能优于其它各组。
The study look forward to establish animal training model by stimulate different altitude training methods through artificial low-pressure oxygen chamber.and apply detection methods of morphological, physiological and biochemical to different training patterns in order to test and analyze the myocardial and blood-related indicators of the rat. Comparative study the influences of morphological and biochemical indicators myocardial cell to the different training methods, and analyze the significance and problems of these methods from the perspective of sport-biochemical and athletic training,to help scientists and coaches understand these issues more comprehensively, meanwhile, make more effective use of these methods in the practice of training.
According to the request of experiment,the big mouse stochastically divided into living low compare group (LoC), livinglow- exerciseing low group (LoLo), Living low-Training high group (LoHi), living-high compare group (HiC), Living high-Training low group (HiLo),high-live-high-exercise group (HiHi), Living high-Exercise high-Training low group (HiHiLo), altogether 7 groups, 10 of each group.With A. X.Bigard method,by ways of increases progressively and great intensity runs training method to simulate low oxygen training model. After gave corresponding simulation plateau training (4 weeks altogether), extracted one big mice out of each group to execute head cut stochastically, took the tissue of cardiac apex, made slice in the thickness of 50nm, observed the changes of the mice myocardium cell on five aspects of the cell nucleus, the mitochondrial, the myofibril, the myocardium intercalated plate and the capillary during the process of the low oxygen training by transmission electron microscopy of JEM2-1230.After the simulation plateau training of other mouse of each group completed, made them running out of strength disposablly, recorded the time, then executed head cut, stripped the heart fast, took the blood sample ,separated blood serum, frozen the tissue of myocardium and the blood serum with liquid nitrogen, preserves by refrigerator(-40℃). Examined the activeness and the content of SOD, LDH, LA, CK, SDH,ATP enzyme in myocardium tissue and blood serum with the reagent box.
The experiment results showed that:
1) Different effects on heart mitochondria appearanced according to different training methods after 4 weeks of simulated hypoxic training, among which LoC, LoLo, HiLo groups are rich in the number of mitochondria, integrity of structural without damages; however, the morphological structure of mitochondrion showed some damages in the LoHi, Hic, HiHi and HiHiLo groups, which performance of unequal size, different shapes, swelling, gathered ridge and reduced number.
2) The influence of myofibrils and muscle cells: In LoC, LoLo, LoHi and HiLo groups, the structural of myofibrils were integrated, the arrangement of myofilaments are neatly and regularly, the sarcomeres were visible clearly and the sarcolemma was intact. In HiC, HiHi, HiHiLo groups, the myofibrils had changes, the arrangements were irregularly with an uneven length, and some local myofilaments lost.
3) The influence of capillary vessel and myocardium intercalated disc: HiHiLo group has some capillary vessel structural damages. in different altitude training. Behaviour destroys for blood vessel bast cell nucleus chromatin aggregation, swelling expand, the bast cell has. The LoHi group myocardium intercalated disc has changes , shows being that the bridge granule falls off , the gap enhances , linking clearance leakage and so on. But, every group capillary vessel and myocardium intercalated disc structure have no obvious loss other without damages.
4) In the view of changes of LA and LDH in serum, different methods of hypoxia training can improve the anaerobic metabolize ablity of myocardium differently, and the training method used by HiLo, LoHi and HiHi groups could improve significantly.
5) In the view of the changes of myocardial energy metabolize indicators, ATP enzyme among the different hypoxia training methods HiLo, HiHiLo and LoHi groups improved the myocardial energy metabolism significantly.
6) In the view of changes of CK and LDH in myocardium, different methods of hypoxia training can improve the anaerobic metabolize ablity of myocardium differently, and the training method used by HiLo, HiHiLo and LoHi groups could improve significantly.
7) The activity of myocardial SOD by HiLo ,LoHi group increased mostly ;The groups which improved the activity of myocardial oxygen metabolism enzyme SDH most significantly is HiLo, LoHi, which suggest that among methods of simulating altitude training, HiLo, LoHi groups can improve the antioxidant capacity and myocardium aerobic metabolism ability of movement rat.
Conclusion:
1) Different model of simulation altitude training have some effects on myocardial ultrastructure with 4 wk training, adaptive changes of myocardial, some grops have improved in function of microvascular, but other groups have some gameges. suggesting that different altitude training may lead to different changes in myocardial ultrastructure.
2) Different altitude training methods have effect on myocardial physiological and biochemical indicators differently, but overall, altitude training can improve body function effectively, improve the capacity of exercise, among different training methods the high-living low-exercise is especially significant.
按实验要求将大鼠随机分为低住安静组(LoC)、低住低练组(LoLo)、低住高练组(LoHi)、高住安静组(HiC)、高住低练组(HiLo)、高住高练组(HiHi),高住高练低训组(HiHiLo),共7组,每组10只。以A.X.Bigard跑台训练方法进行递增、大强度的跑台训练方式建立模拟低氧训练模型。各组大鼠在给予相应的模拟高原训练后(共4周),每组随机抽取一只断头处死,取心尖组织,制成50nm厚度切片,采用JEM2-1230型透射电镜技术从细胞核、线粒体、肌原纤维、心肌闰盘和毛细血管等五个方面观察大鼠心肌细胞超微结构在低氧训练过程中的变化。其余各组大鼠在模拟高原训练完成后,一次性在跑台跑至力竭,记录力竭时间,后断头处死,快速剥离心脏,取血液样品分离血清,将心肌组织和血清液氮冷冻,后置于冰箱(-40℃)保存。用试剂盒检测心肌组织和血清中的SOD、LDH、LA、CK、SDH、ATP酶的活性和含量。
实验结果表明:
1)对心肌线粒体的影响,其中LoC、LoLo、HiLo组的线粒体数目丰富、个数较多,结构完整无损伤;而LoHi、HiC、HiHi、HiHiLo组线粒体形态结构有不同程度的损伤,表现为体积大小不均,形态各异,有肿胀,嵴聚集,数目减少等。
2)对肌原纤维和肌细胞核的影响,LoC、LoLo、LoHi、HiLo组肌原纤维和肌细胞核结构完整,肌丝排列规则整齐,肌节清晰可见,肌膜完整;HiC、HiHi、HiHiLo组肌原纤维有变化,排列不整齐,长短不均一,部分局域肌丝缺失。
3)对心肌微血管和闰盘的影响,不同高原训练方式中HiHiLo组微血管有部分结构损伤,表现为血管内皮细胞核染色质聚集,肿胀膨大,内皮细胞有破坏。LoHi组心肌闰盘有变化,表现为桥粒减少、间隙增大、连接不紧密等。
4)对血清LA和LDH的影响,不同的低氧训练方法中对大鼠无氧代谢能力有不同的改善作用,其中以HiLo、LoHi和HiHi组值低于其他各训练组,提示其对乳酸有较好的耐受和清除能力。
5)对心肌ATP酶的影响,Na~+-K~+ATP酶以HiHiLo、HiLo组;Ca~(2+)、Mg~(2+)-ATPase以HiLo、HiHiLo、LoHi组酶活性优于其它各组。提示,主要的模拟高原训练方法HiLo、HiHiLo和LoHi组对心肌的ATP酶活性都有比较显著的改善作用。
6)心肌LDH值以HiHiLo组较高,心肌CK以HiLo、LoHi、HiHiLo组活性高于其他各组。提示,不同的低氧训练方法中对心肌无氧代谢改善作用比较显著的训练方法是HiLo、HiHiLo和LoHi组。
7)HiLo、LoHi法对心肌SOD活性的提高较其它各组显著。心肌有氧代谢酶SDH活性提高较显著的组是HiLo、LoHi组,提示模拟高原训练方法HiLo、LoHi训练法对运动大鼠心肌抗氧化能力和有氧代谢能力有较好的改善作用。。
结论:
1)经过4wk模拟不同高原训练,对心肌超微结构有不同的影响,HiLo组LoLo组心肌超微结构有适应性变化,而HiHiLo、HiHi组心肌结构在线粒体、微血管、肌纤维、心肌闰盘等方面表现有不同程度的损伤。提示,模拟高原训练方法对心肌超微结构的影响因训练方法的异同而产生不同的效果。
2)不同高原训练方式对心肌生理生化指标有不同影响,但整体来看,高原训练可以有效的改善机体机能,提高运动能力,不同的训练方法中尤以高住低练法较为显著,可能优于其它各组。
The study look forward to establish animal training model by stimulate different altitude training methods through artificial low-pressure oxygen chamber.and apply detection methods of morphological, physiological and biochemical to different training patterns in order to test and analyze the myocardial and blood-related indicators of the rat. Comparative study the influences of morphological and biochemical indicators myocardial cell to the different training methods, and analyze the significance and problems of these methods from the perspective of sport-biochemical and athletic training,to help scientists and coaches understand these issues more comprehensively, meanwhile, make more effective use of these methods in the practice of training.
According to the request of experiment,the big mouse stochastically divided into living low compare group (LoC), livinglow- exerciseing low group (LoLo), Living low-Training high group (LoHi), living-high compare group (HiC), Living high-Training low group (HiLo),high-live-high-exercise group (HiHi), Living high-Exercise high-Training low group (HiHiLo), altogether 7 groups, 10 of each group.With A. X.Bigard method,by ways of increases progressively and great intensity runs training method to simulate low oxygen training model. After gave corresponding simulation plateau training (4 weeks altogether), extracted one big mice out of each group to execute head cut stochastically, took the tissue of cardiac apex, made slice in the thickness of 50nm, observed the changes of the mice myocardium cell on five aspects of the cell nucleus, the mitochondrial, the myofibril, the myocardium intercalated plate and the capillary during the process of the low oxygen training by transmission electron microscopy of JEM2-1230.After the simulation plateau training of other mouse of each group completed, made them running out of strength disposablly, recorded the time, then executed head cut, stripped the heart fast, took the blood sample ,separated blood serum, frozen the tissue of myocardium and the blood serum with liquid nitrogen, preserves by refrigerator(-40℃). Examined the activeness and the content of SOD, LDH, LA, CK, SDH,ATP enzyme in myocardium tissue and blood serum with the reagent box.
The experiment results showed that:
1) Different effects on heart mitochondria appearanced according to different training methods after 4 weeks of simulated hypoxic training, among which LoC, LoLo, HiLo groups are rich in the number of mitochondria, integrity of structural without damages; however, the morphological structure of mitochondrion showed some damages in the LoHi, Hic, HiHi and HiHiLo groups, which performance of unequal size, different shapes, swelling, gathered ridge and reduced number.
2) The influence of myofibrils and muscle cells: In LoC, LoLo, LoHi and HiLo groups, the structural of myofibrils were integrated, the arrangement of myofilaments are neatly and regularly, the sarcomeres were visible clearly and the sarcolemma was intact. In HiC, HiHi, HiHiLo groups, the myofibrils had changes, the arrangements were irregularly with an uneven length, and some local myofilaments lost.
3) The influence of capillary vessel and myocardium intercalated disc: HiHiLo group has some capillary vessel structural damages. in different altitude training. Behaviour destroys for blood vessel bast cell nucleus chromatin aggregation, swelling expand, the bast cell has. The LoHi group myocardium intercalated disc has changes , shows being that the bridge granule falls off , the gap enhances , linking clearance leakage and so on. But, every group capillary vessel and myocardium intercalated disc structure have no obvious loss other without damages.
4) In the view of changes of LA and LDH in serum, different methods of hypoxia training can improve the anaerobic metabolize ablity of myocardium differently, and the training method used by HiLo, LoHi and HiHi groups could improve significantly.
5) In the view of the changes of myocardial energy metabolize indicators, ATP enzyme among the different hypoxia training methods HiLo, HiHiLo and LoHi groups improved the myocardial energy metabolism significantly.
6) In the view of changes of CK and LDH in myocardium, different methods of hypoxia training can improve the anaerobic metabolize ablity of myocardium differently, and the training method used by HiLo, HiHiLo and LoHi groups could improve significantly.
7) The activity of myocardial SOD by HiLo ,LoHi group increased mostly ;The groups which improved the activity of myocardial oxygen metabolism enzyme SDH most significantly is HiLo, LoHi, which suggest that among methods of simulating altitude training, HiLo, LoHi groups can improve the antioxidant capacity and myocardium aerobic metabolism ability of movement rat.
Conclusion:
1) Different model of simulation altitude training have some effects on myocardial ultrastructure with 4 wk training, adaptive changes of myocardial, some grops have improved in function of microvascular, but other groups have some gameges. suggesting that different altitude training may lead to different changes in myocardial ultrastructure.
2) Different altitude training methods have effect on myocardial physiological and biochemical indicators differently, but overall, altitude training can improve body function effectively, improve the capacity of exercise, among different training methods the high-living low-exercise is especially significant.
引文
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