间歇性低氧训练对红细胞生成及其抗氧化能力的影响
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摘要
高原训练一直以来被认为是提高有氧运动能力的训练手段，并被广泛应用到各种运动项目中。但在高原训练时运动强度难以提高，为了避免这个不利因素，人们提出了各种低氧训练方法来代替高原训练，间歇性低氧训练就是一种新兴的模拟高原训练的方法。本文观察了4周的间歇性低氧训练对人体血液中红细胞生成及其抗氧化能力的影响，以及测试运动能力相关指标，试想分析间歇性低氧训练对提高人体最大吸氧量和有氧运动能力有积极作用和显著效果，为间歇性低氧训练在运动训练中的可行性提供实验依据。
实验方法：
本实验将华南师范大学2000级体育系健康男生15名随机分为两组，对照组，间歇性吸低氧组。间歇性低氧组：每天和对照组一样的正常身体活动，另外在安静状态下间歇性吸低氧总共50-60分钟，6天／周，连续4周，氧浓度14％、12％、10％，逐渐降低。两组分别在低氧训练前、低氧训练的第1、3、10、15、17、22、24天，低氧训练结束后的第5、8、14天抽取静脉血2ml，用于测定红细胞参数，网织红细胞参数，促红细胞生成素，可溶性转铁蛋白受体；分别在低氧训练前一天、低氧训练结束后一天采用Bruce递增负荷运动至力竭的方案在跑台上运动，并在安静时、力竭运动后3分钟内取静脉血3ml，用于测定红细胞自由基和抗氧化系统指标；同时测定血乳酸、心率、最大吸氧量、通气量、运动到达力竭的时间。
实验结果：
(1)间歇性低氧训练可以促进EPO的分泌，引起造血系统的活跃；EPO是在低氧刺激后一天就升高，如果低氧浓度不变，在达到峰值后即开始降低；间歇性低氧训练引起的sTfr升高是在EPO升高一周后开始显著升高，并在停止低氧训练后15天仍显著高于正常水平，提示间歇性低氧训练过程中和停止后，均有红细胞和血红蛋白的生成增多的可能。
卢币q店义伞q才存伞伞魔＊口o三忐吩士乒仁1文 叨不友
（2）间歇性低氧训练可以引起红细胞计数和压积明显升高，说明本实验所采用地
间歇性低氧训练可以取得与其它形式低氧训练一样的效果。鉴于本实验Hb浓度没
有变化，希望在以后的研究中，测量血容量或血红蛋白总量，并在营养上给予一定
的监控，进一步探讨间歇性低氧训练促进血红蛋白的合成情况。
（3）Bruce递增负荷的力竭运动可引起红细胞中 MDA生成显著增多，GSH干X酶活
性明显增加，而SOD酶活性和HRPRP含量变化不大，说明Bruce方法的递增负荷的
力竭运动导致红细胞自山基生成增多，可引起红细胞氧化应激损伤；GSH干X酶、
SOD酶活性和HRPRP在运动后的不同变化与本实验采用的运动方式有关。
（4）间歇性低氧训练可提高机体抗氧化能力，4周的低氧训练后，红细胞抗氧化酶
SOD、GSH干X活性和 HRPRP的含量在安静时和力竭运动后都得到明显提高，说明间
歇性低氧刺激机体生成自由基增多，诱导抗氧化酶和HRPRP生成增多以抵御自由基
的对红细胞地侵害。提示可以采用间歇性低氧训练来增强红细胞抗氧化的能力，有
助于改善有氧运动能力。
（5）间歇性低氧训练可提高机体最大吸氧量，延长力竭运动时间。说明间歇性低
氧训练可提高有氧运动能力。
本实验通过分析间歇性低氧训练可增加红细胞生成和抗氧化能力，提供改善有
氧运动能力的机制上的实验依据；通过分析间歇性低氧训练可提高最大吸氧量和有
氧运动能力为间歇性低氧训练在运动训练中应用地有效性和可行性提供实验依据。
Training at moderate altitude is often used by athletes from a variety of sports to improve their aerobic capacity. In order to avoid the detrimental effects of reduced training intensity at hypoxic condition, many simulated altitude training have been supposed. Intermittent normobaric hypoxia training is one style of simulated altitude training too.
The study was to investigate that effects of 4 weeks intermittent hypoxia training(IHT) on erythropoiesis and the antioxidant ability in erythrocytes. Some markers in realation to erythorpoiesis and antioxidant system were measured and analyzed. We undertook these data in order to test the hypothesis that IHT is helpful to exercise performance and maximal oxygen uptake (Vo2max).
Our research subjects are 15 male students who were randomly divided into groups of control and of IHT. Two groups took part in same daily study and exercise during the experiment. The INT group inhaled a intermittent hypoxia gas mixture (O2% is 14%, 12% , 10%) 50-60 minutes daily for 4 weeks (6d/week). Blood of two groups were collected 2ml for measuring red cell count, hematocrit (Hct), hemoglobin (Hb), reticulocytes hematocrit (HctRet), erythropoietin (EPO) and soluable serum transferrin receptor (sTfr) before IHT, on days 1, 3, 10, 15, 17, 22, 24 during IHT and on days 5, 8,14 of post-IHT. Both groups completed incremental exercise to exhaustion on the treadmill before and after IHT. The MDA and human red cell protect protein (HRPRP) concentration, SOD and GSH-PX activity and blood lactic acid were measured at rest and post-exercise respectively. The Voimax , heard rate (HR) and the time to exhaustion were measured as well.
The results are follow:
1. In our study, IHT could stimulate erythropoietin secretion that lead to erythpopoietic response. We observed that serum EPO significantly increased after one day hypoxia. After the initial peak, serum EPO decreased if the concentration of oxygen was not decreased continually. The measurement of sTfr was used as indicator of erythropoietic activity as EPO. In our study, the sTfr show a significantly increased after EPO has been increased for one week and still remained high level after IHT. It reflect an activity of erythropoiesis increase both duing IHT and post-IHT.
2. In our study, the red cell count and Hct were significantly elevated since IHT. It indicate that intermittent exposure to hypoxia 50-60 minutes daily for 4 weeks induce a similar stimulation of erythropoiesis as high- altitude-training. The Hb values did not show any significant changes during our experiment. We suggest that the blood volume and total hemoglobin mass should be measured and nutrition supply should be monitored and controled in later research of IHT.
3. The MDA concentration and activity of GSH-PX in erythrocytes were significantly increased after incremental exercise to exhaustion. At the same time, activity of SOD and the concentration of HRPRP in erythrocytes did not change significantly. It indicate that the possibility of oxidative damage in erythrocytes was caused by reactive oxygen species (ROS) increasing post-exercise. The different showing of GSH-PX, SOD activities and HRPRP may be relate to the type of exercise what we took.
4. In our study, antioxidant capacity showed a statistically increasing post-hypoxia compared with pre-hypoxia. The activities of SOD, GSH-PX and concentration of HRPRP were increased both at rest and post-exercise compared with what they were 4 weeks before. It show that hypoxia increased ROS which caused elevating of activity of antioxidant enzymes and concentration of HRPRP. It suggested that INT could increase antioxidant capacity and be helpful to improve arobic capacity.
5. This study show that Vo2max increased and the time to exhaustion decreased significantly after 4 weeks of IHT. It indicated that sports performance can be enhanced effectively by IHT. It suggested that IHT may be used in the sport training.
实验方法：
本实验将华南师范大学2000级体育系健康男生15名随机分为两组，对照组，间歇性吸低氧组。间歇性低氧组：每天和对照组一样的正常身体活动，另外在安静状态下间歇性吸低氧总共50-60分钟，6天／周，连续4周，氧浓度14％、12％、10％，逐渐降低。两组分别在低氧训练前、低氧训练的第1、3、10、15、17、22、24天，低氧训练结束后的第5、8、14天抽取静脉血2ml，用于测定红细胞参数，网织红细胞参数，促红细胞生成素，可溶性转铁蛋白受体；分别在低氧训练前一天、低氧训练结束后一天采用Bruce递增负荷运动至力竭的方案在跑台上运动，并在安静时、力竭运动后3分钟内取静脉血3ml，用于测定红细胞自由基和抗氧化系统指标；同时测定血乳酸、心率、最大吸氧量、通气量、运动到达力竭的时间。
实验结果：
(1)间歇性低氧训练可以促进EPO的分泌，引起造血系统的活跃；EPO是在低氧刺激后一天就升高，如果低氧浓度不变，在达到峰值后即开始降低；间歇性低氧训练引起的sTfr升高是在EPO升高一周后开始显著升高，并在停止低氧训练后15天仍显著高于正常水平，提示间歇性低氧训练过程中和停止后，均有红细胞和血红蛋白的生成增多的可能。
卢币q店义伞q才存伞伞魔＊口o三忐吩士乒仁1文 叨不友
（2）间歇性低氧训练可以引起红细胞计数和压积明显升高，说明本实验所采用地
间歇性低氧训练可以取得与其它形式低氧训练一样的效果。鉴于本实验Hb浓度没
有变化，希望在以后的研究中，测量血容量或血红蛋白总量，并在营养上给予一定
的监控，进一步探讨间歇性低氧训练促进血红蛋白的合成情况。
（3）Bruce递增负荷的力竭运动可引起红细胞中 MDA生成显著增多，GSH干X酶活
性明显增加，而SOD酶活性和HRPRP含量变化不大，说明Bruce方法的递增负荷的
力竭运动导致红细胞自山基生成增多，可引起红细胞氧化应激损伤；GSH干X酶、
SOD酶活性和HRPRP在运动后的不同变化与本实验采用的运动方式有关。
（4）间歇性低氧训练可提高机体抗氧化能力，4周的低氧训练后，红细胞抗氧化酶
SOD、GSH干X活性和 HRPRP的含量在安静时和力竭运动后都得到明显提高，说明间
歇性低氧刺激机体生成自由基增多，诱导抗氧化酶和HRPRP生成增多以抵御自由基
的对红细胞地侵害。提示可以采用间歇性低氧训练来增强红细胞抗氧化的能力，有
助于改善有氧运动能力。
（5）间歇性低氧训练可提高机体最大吸氧量，延长力竭运动时间。说明间歇性低
氧训练可提高有氧运动能力。
本实验通过分析间歇性低氧训练可增加红细胞生成和抗氧化能力，提供改善有
氧运动能力的机制上的实验依据；通过分析间歇性低氧训练可提高最大吸氧量和有
氧运动能力为间歇性低氧训练在运动训练中应用地有效性和可行性提供实验依据。
Training at moderate altitude is often used by athletes from a variety of sports to improve their aerobic capacity. In order to avoid the detrimental effects of reduced training intensity at hypoxic condition, many simulated altitude training have been supposed. Intermittent normobaric hypoxia training is one style of simulated altitude training too.
The study was to investigate that effects of 4 weeks intermittent hypoxia training(IHT) on erythropoiesis and the antioxidant ability in erythrocytes. Some markers in realation to erythorpoiesis and antioxidant system were measured and analyzed. We undertook these data in order to test the hypothesis that IHT is helpful to exercise performance and maximal oxygen uptake (Vo2max).
Our research subjects are 15 male students who were randomly divided into groups of control and of IHT. Two groups took part in same daily study and exercise during the experiment. The INT group inhaled a intermittent hypoxia gas mixture (O2% is 14%, 12% , 10%) 50-60 minutes daily for 4 weeks (6d/week). Blood of two groups were collected 2ml for measuring red cell count, hematocrit (Hct), hemoglobin (Hb), reticulocytes hematocrit (HctRet), erythropoietin (EPO) and soluable serum transferrin receptor (sTfr) before IHT, on days 1, 3, 10, 15, 17, 22, 24 during IHT and on days 5, 8,14 of post-IHT. Both groups completed incremental exercise to exhaustion on the treadmill before and after IHT. The MDA and human red cell protect protein (HRPRP) concentration, SOD and GSH-PX activity and blood lactic acid were measured at rest and post-exercise respectively. The Voimax , heard rate (HR) and the time to exhaustion were measured as well.
The results are follow:
1. In our study, IHT could stimulate erythropoietin secretion that lead to erythpopoietic response. We observed that serum EPO significantly increased after one day hypoxia. After the initial peak, serum EPO decreased if the concentration of oxygen was not decreased continually. The measurement of sTfr was used as indicator of erythropoietic activity as EPO. In our study, the sTfr show a significantly increased after EPO has been increased for one week and still remained high level after IHT. It reflect an activity of erythropoiesis increase both duing IHT and post-IHT.
2. In our study, the red cell count and Hct were significantly elevated since IHT. It indicate that intermittent exposure to hypoxia 50-60 minutes daily for 4 weeks induce a similar stimulation of erythropoiesis as high- altitude-training. The Hb values did not show any significant changes during our experiment. We suggest that the blood volume and total hemoglobin mass should be measured and nutrition supply should be monitored and controled in later research of IHT.
3. The MDA concentration and activity of GSH-PX in erythrocytes were significantly increased after incremental exercise to exhaustion. At the same time, activity of SOD and the concentration of HRPRP in erythrocytes did not change significantly. It indicate that the possibility of oxidative damage in erythrocytes was caused by reactive oxygen species (ROS) increasing post-exercise. The different showing of GSH-PX, SOD activities and HRPRP may be relate to the type of exercise what we took.
4. In our study, antioxidant capacity showed a statistically increasing post-hypoxia compared with pre-hypoxia. The activities of SOD, GSH-PX and concentration of HRPRP were increased both at rest and post-exercise compared with what they were 4 weeks before. It show that hypoxia increased ROS which caused elevating of activity of antioxidant enzymes and concentration of HRPRP. It suggested that INT could increase antioxidant capacity and be helpful to improve arobic capacity.
5. This study show that Vo2max increased and the time to exhaustion decreased significantly after 4 weeks of IHT. It indicated that sports performance can be enhanced effectively by IHT. It suggested that IHT may be used in the sport training.
引文
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54．王茂叶，雷志平．间歇性低氧训练对小鼠有氧代谢能力影响的研究．西安体育学院学报．2001，18(1)：32-34
55. Bigard AX, et al. Skeletal muscle changes after endurance training at high altitude. J Appl Phsiol. 1991,71(6):2114-2121
56．黄庆愿，高钰琪，孙秉庸．缺氧及缺氧复合运动大鼠骨骼肌组织化学观察．中国运动医学杂志．2000，19(4)：204-206
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61. Mizuno M, et al. Limb skeletal muscle adaptation in athletes after training at altitude. J Appl Physiol. 1990,68(2):496-502
62．李世成，田野．模拟高原训练对小鼠骨骼肌LDH同工酶谱的影响．武汉体育学院学报．2000，3：
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79. Sawka MN. et al. Altitude acclimatization and blood wolume:effects of exogenous erythrocyte volume expansion. J Appl Physiol. 1996,81:636-642
80. Bning D, et al. After-effects of a high altitude expendition on blood. Int J Sports Med. 1997, 18:179-185
81. Robach P, et al. Operation everest Ⅲ: role of plasma volume expansion on VO2max during prolonged high-altitude exposure. J Appl Physiol. 2000, 89:29-37
82. Chapman RF, Stray-Gundersen and Levine BD. Individual variation in altitude training. J Appl Physiol. 1998, 85, 1448-1455
83．李晖等．递增负荷力竭性运动时大鼠血液氧化、抗氧化能力及RBCM生物物理特性得研究．中国运动医学杂志．2001，20(3)：256-259
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85．张爱芳，王贵，胡艳龙．一次性递增负荷运动对足球运动员红细胞抗氧化能力和Na+_K+_ATP酶活性的影响．中国运动医学杂志．2000，19(4)：429-431
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87. Chao WH, et al. Oxidative stress in humans during work at moderate altitude. J Nutr. 1999, 129(11) :2009-2012
88．周兆年等．间歇性低氧对心脏的保护作用．中国应用生理学杂志．2000，16(3)：34-36
89．张缨，文茹．运动性贫血的发生机制与监测．北京体育大学学报．2001，24(3)：331-334
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53. Gundersen JS, Chapman RF and Levine BD. "Living high-training low" altitude training improves sea level performance in male and female elite runners. J Appl Physiol. 2001,91:1113-1120
54．王茂叶，雷志平．间歇性低氧训练对小鼠有氧代谢能力影响的研究．西安体育学院学报．2001，18(1)：32-34
55. Bigard AX, et al. Skeletal muscle changes after endurance training at high altitude. J Appl Phsiol. 1991,71(6):2114-2121
56．黄庆愿，高钰琪，孙秉庸．缺氧及缺氧复合运动大鼠骨骼肌组织化学观察．中国运动医学杂志．2000，19(4)：204-206
57. Desplanches D, Hoppeler H, Tuscher L. Muscle tissue adaptations to training in normoxia hypoxia. J Appl Physiol. 1996,81(5):1946-1951
58. Terrados N, et al. Is hypoxia a stimulus for synthesis of oxidative enzymes and myoglobin? J Appl Physiol. 1990,68(6):2369-2373
59. Terrodos N, Altitude training and muscle metabolism. Int J Sports Med. 1992,13(1):206-208
60. Green H, et al. Downregulation of Na~+-K~+-ATPase pumps in skeletal muscle with training in normobaric hypoxia. J Appl Physiol. 1999,86(5):1745-1748
61. Mizuno M, et al. Limb skeletal muscle adaptation in athletes after training at altitude. J Appl Physiol. 1990,68(2):496-502
62．李世成，田野．模拟高原训练对小鼠骨骼肌LDH同工酶谱的影响．武汉体育学院学报．2000，3：
63. Melissa L, et al. Skeletal muscle adaptation to training under normobaric hypoxic versus normoxic conditions. Med Sci Sports Exerc. 1997,29:238-243
64. Clanton TL, Klawitter PF. Physiological and genomic consequences of intermittent hypoxia invited review: Adaptive responses of skeletal muscle to intermittent hypoxia: the know and the unknown. J Appl Physiol. 2001,90:2476-2487
65. Clanton TL, et al. Improved anoxic tolerance in rat diaphragm following intermittent hypoxia. J Appl Physiol. 2001,90:2508-2013
66. Robinson DM, et al. Training intensity of elite male distance runners. Med Sci Sports Exerc, 1991,23:1078-1082
67. Chapman RF, Michael E and Stager JM. Degree of arterial desaturation in normoxia influences VO2max decline in mild hypoxia. Med Sci Sports Exerc. 1999,31(5):658-663
68. Gavin TP, Stager JM and Derchak PA. Ventilation' s role in the decline in VO2max
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69．雷志平等．间歇性低氧训练对气体代谢及乳酸阈影响的实验研究．六科大汇编(二)．
70. Parisotto R, et al. A novel method utilizing markers of altered erythropoiesis for the detection of recombinant human erythropoietin abuse in athletes. Haematologica. 2000, 85 (6): 564-572
71. Schmidt W, et al. Effects of maximal and submaximal exercise under normoxic and hypoxic conditions on serum erythropietin level. Int J Sports Med. 1991, 12:457-461
72. Knaupp W, et al. Erythropietin response to acute normobaric hypoxia in humans. J Appl Physiol. 1992, 73:837-840
73．冯连世等．高原训练对中长跑运动员红细胞生成的作用．体育科学．1998，18(4)：78-81
74. Birkeland KI, et al. Effect of rhEPO administration on serum levels of sTfr and cycling performance. Med Sci Sports Exerc. 2000, 32 (7): 1238-1243
75．冯连世．高原训练及其研究现状．体育科学．1999，19(6)：66-71
76. Wolfer EE, et al. Oxygen transport during steady stade submaximal exercise in chronic hypoxia. J Appl Physiol. 1991, 70:1129-1136
77. Stray-Gundersen J, Hochstein A and Vebine BD. Effect of 4 weeks altitude training exposure and training on red cell mass in trained runners. Med Sci Sports Exerc. 1992, 24(suppl): S90
78. Friedmann B, et al. Effects of iron supplementation on total hemoglobin during endurance training at moderate altitude. Int J Sports Med. 1999,20:78-85
79. Sawka MN. et al. Altitude acclimatization and blood wolume:effects of exogenous erythrocyte volume expansion. J Appl Physiol. 1996,81:636-642
80. Bning D, et al. After-effects of a high altitude expendition on blood. Int J Sports Med. 1997, 18:179-185
81. Robach P, et al. Operation everest Ⅲ: role of plasma volume expansion on VO2max during prolonged high-altitude exposure. J Appl Physiol. 2000, 89:29-37
82. Chapman RF, Stray-Gundersen and Levine BD. Individual variation in altitude training. J Appl Physiol. 1998, 85, 1448-1455
83．李晖等．递增负荷力竭性运动时大鼠血液氧化、抗氧化能力及RBCM生物物理特性得研究．中国运动医学杂志．2001，20(3)：256-259
84．辛东等．力竭性运动时大鼠脑组织自由基产生及氧化、抗氧化能力得动态观察．中国运动医学杂志．1999，18(4)：321-323
85．张爱芳，王贵，胡艳龙．一次性递增负荷运动对足球运动员红细胞抗氧化能力和Na+_K+_ATP酶活性的影响．中国运动医学杂志．2000，19(4)：429-431
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86．张之南，李蓉生．红细胞疾病基础与临床．北京．科学出版社．2000
87. Chao WH, et al. Oxidative stress in humans during work at moderate altitude. J Nutr. 1999, 129(11) :2009-2012
88．周兆年等．间歇性低氧对心脏的保护作用．中国应用生理学杂志．2000，16(3)：34-36
89．张缨，文茹．运动性贫血的发生机制与监测．北京体育大学学报．2001，24(3)：331-334
90．陈文鹤，魏安奎．红细胞变形性对运动能力的影响．上海体育学院学报．1995，19(3)：69-72
91．吴歆，顾熊飞．人红细胞抗氧化蛋白的抗氧化机制及对亚急性衰老小鼠保护作用的初步研究．中山医科大学硕士研究生学位论文．2000，6
92．胡扬，黄亚茹．耐力训练的新方法—高住低训法．体育科学．2001，21(2)：66-70