高原低氧环境大强度运动后增压辅助方法对大鼠骨骼肌中HIF-1α及骨骼肌能量代谢的影响

发布时间：2018-02-28 08:48

本文关键词： 高原低氧增压辅助大鼠骨骼肌 HIF-1α 能量代谢　出处：《青海师范大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：通过建立大鼠在高原低氧环境下大强度运动后，将其转移至增压舱内在0.2-0.3MPa（绝对压力2-3ATA）的压力环境中恢复的动物模型，探讨高原低氧环境下大强度训练与施加增压辅助方法对大鼠骨骼肌组织中HIF-1α、GLUT-1、GLUT-4蛋白表达及LDH、SDH酶活性的变化对大鼠骨骼肌疲劳恢复的影响。方法：健康雄性Wistar大鼠56只随机分为7组，每组8只：其中A组为安静对照组，B组为自然恢复组，C组为0.2MPa增压1h恢复组，D组0.2MPa增压2h恢复组，E组为0.3MPa增压1h恢复组，F组为低氧运动后自然恢复组，G组为低氧运动后0.2MPa增压1h恢复组；平均体重A组197.71±16.87g，B组210.25±12.06g，C组208.38±4.75g，D组208.00±14.33g，E组210.88±11.99g，F组212.75±12.09g，G组210.50±8.53g。7组大鼠在西宁（2260m）青海大学医学院动物实验房进行3天适应性训练和6天正式训练:其中适应期第1天跑速15m/min，其后每天递增5m/min；适应期坡度为0°；运动时间第一天15min，第二天20min，第三天30min。正式训练第1天跑速25m/min，第2天30m/min，以后4天维持此跑速至实验结束；第1，2天坡度0°第3天增至10°，余下3天维持此坡度不变；正式训练期间运动时间60min/天。第7天进行B组至G组速度为35m/mim、坡度为10°的力竭运动。运动后依照力竭运动组别顺序除B组、F组外其余各组进舱增压恢复休息1h,增压恢复结束后24h所有大鼠实施腹腔麻醉取一侧腓肠肌，运用蛋白质免疫印迹法检测各组HIF-1α、GLUT-1、GLUT-4的蛋白表达，运用ELISA法检测各组LDH和SDH酶活性。结果：（1）通过7天高原低氧环境下大强度运动后施加增压辅助方法的恢复实验发现，大鼠骨骼肌HIF-1α蛋白表达呈现先下降后上升再下降的趋势，其中C组（0.2MPa增压1h恢复）上升幅度较大。（2）通过7天高原低氧环境下大强度运动后施加增压辅助方法的恢复实验发现，大鼠骨骼肌GLUT-1蛋白表达呈现先上升后下降再上升的趋势，其中G组（低氧运动后0.2MPa的增压1h恢复）上升较大。（3）通过7天高原低氧环境下大强度运动后施加增压辅助方法的恢复实验发现，大鼠骨骼肌GLUT-4蛋白表达呈现先下降后上升的趋势，其中G组（低氧运动后0.2MPa的增压1h恢复）上涨幅度最大。 (4)通过7天增压辅助方法干预后，G组（低氧运动后增压恢复）与F组（低氧运动后自然恢复）相比大鼠骨骼肌LDH活性上升且两组具有显著性差异（P0.05） (5)通过7天增压辅助方法干预后，大鼠骨骼肌SDH活性从A组（安静对照）至G组（低氧运动后增压恢复）呈现先上升后下降再上升再下降的趋势且各组之间的差异不具有显著性。结论：（1）增压辅助方法干预的C组（0.2MPa增压1h恢复）、D组（0.2MPa增压2h恢复）、E组（0.3MPa增压1h恢复）大鼠骨骼肌HIF-1α蛋白表达量上调，表明施加增压辅助方法对大鼠骨骼肌HIF-1α产生影响，HIF-1α表达上升可诱导其下游的靶基因转录上调增加骨骼肌氧及营养物质的供应，有利于高原低氧环境下大强度训练后的疲劳恢复。（2）增压辅助干预后G组（低氧运动后0.2MPa的增压1h恢复）与F组（低氧运动后自然恢复）相比GLUT-1、GLUT-4有明显上升趋势。GLUT-1、GLUT-4表达上升可增加骨骼肌的葡萄糖转运能力，有利于骨骼肌对葡萄糖的摄取及能量物质的补充，促进机体在高原低氧环境下大强度训练后的疲劳恢复。（3）增压辅助干预后C组（0.2MPa增压1h恢复），E组（0.3MPa增压1h恢复）大鼠骨骼肌LDH的活性上调；G组（低氧运动后0.2MPa增压1h恢复）与F组（低氧运动后自然恢复）相比具有显著性（P0.05）。表明增压辅助方法干预的恢复组LDH活性上调可促进骨骼肌乳酸至丙酮酸的代谢，此过程可减少骨骼肌中乳酸的含量，增加对骨骼肌能量的供应，有助于高原低氧环境运动后机体的疲劳恢复。（4）增压辅助干预后大鼠骨骼肌SDH活性无显变化，表明高原低氧大强度运动后采用增压辅助方法对提高大鼠有氧代谢能力的影响不大。
[Abstract]:Objective: to establish rat in high intensity exercise at high altitude, transferred to the pressurized cabin inner 0.2-0.3MPa (absolute pressure 2-3ATA) animal model of restoring pressure in the environment, explore the plateau environment of high intensity training and applied pressure assisted method for HIF-1 rat skeletal muscle alpha, GLUT-1, GLUT-4 the expression of LDH protein and enzyme activity, influence of SDH on the recovery of skeletal muscle fatigue in rats.
Methods: 56 healthy male Wistar rats were randomly divided into 7 groups, 8 rats in each group: group A was the control group, B group, C group natural recovery group, recovery group 0.2MPa 1H group D 0.2MPa turbocharged and supercharged 2H recovery group, E group for 0.3MPa turbocharged 1H recovery group F group, hypoxia after exercise the natural recovery group, G group after hypoxic exercise 0.2MPa turbocharged 1H recovery group; the average weight of group A was 197.71 + 16.87g, 210.25 + 12.06g B group, C group and D group was 208.38 + 4.75g, 208 + 14.33g, 210.88 + 11.99g E group, F group of 212.75 + 12.09g, 210.50 + 8.53g.7 group and G group rats in Xining (2260m) real animal experiments in Medical College of Qinghai University for 3 days and 6 days of formal training adaptation training: the adaptation period of first days of running 15m/min, then added every day 5m/min; adaptation period of slope is 0 degrees; the movement time of the first day of 15min, 20min in second days, third days 30min. formal training first days running 25m/min second. 30m/min day, after 4 days The end of the experiment to maintain the running speed; day 1,2 slope 0 degrees to 10 degrees for third days, the remaining 3 day to maintain this invariant gradient; the formal training exercise during the time is 60min/ days. Seventh days B group G and group velocity is 35m/mim, slope of 10 degrees for the exhaustive exercise. After exercise in exhaustive exercise groups the sequence in B group, F group into the cabin pressurization recovery 1H, booster recovery after 24h rats were anesthetized for the implementation of all side of the gastrocnemius muscle were detected by Western blotting of HIF-1 alpha, GLUT-1, expression of GLUT-4 protein, using ELISA method to detect the LDH and SDH activity:
(1) after 7 days of high altitude hypoxic environment, the HIF-1 C protein expression in the skeletal muscle of rats showed a trend of first decline, then increased and then decreased. The C group (0.2MPa booster 1H recovery) increased significantly.
(2) after 7 days of high altitude hypoxic environment, after boosting exercise, the GLUT-1 protein expression in skeletal muscle of rats showed a trend of first rise, then decline and then increased. Among them, G group (0.2MPa booster 1H recovery after hypoxia exercise) increased significantly.
(3) after 7 days of high-altitude hypoxia, the recovery of GLUT-4 protein expression in skeletal muscle of rats showed a trend of first decline and then increased. After that, G group (0.2MPa booster 1H recovery after hypoxia) increased the most.
(4) after 7 days of booster intervention, the activity of LDH in skeletal muscle of G group increased significantly compared with that in group F (after hypoxic exercise), and there was a significant difference between the two groups (P0.05).
(5) after 7 days of pressurization assisted intervention, the SDH activity of skeletal muscle from A group (quiet control) to G group (after hypoxic exercise supercharging recovery) increased at first, then decreased, then increased and then decreased.
(1) C group intervention (0.2MPa booster assisted method, turbocharged 1H recovery) D group (0.2MPa group, E turbocharged 2H recovery (0.3MPa) turbocharged 1H recovery) expression in rat skeletal muscle HIF-1 protein was up-regulated, showed that applying boost auxiliary method influence on HIF-1 alpha in rat skeletal muscle, the expression of HIF-1 a target up regulation of gene transcription induced increased downstream of the increase in skeletal muscle oxygen and nutrient supply, is conducive to the recovery of fatigue after high intensity training in hypoxia.
(2) after the intervention group G (auxiliary booster booster 1H 0.2MPa recovery after hypoxic exercise group (F) and natural recovery after hypoxic exercise) compared to GLUT-1, GLUT-4 has significantly increased.GLUT-1, increased GLUT-4 expression can increase glucose transport in skeletal muscle, which is beneficial to increase in skeletal muscle glucose uptake and energy the material, promote recovery of body fatigue in high intensity training in altitude hypoxia environment.
The auxiliary booster (3) after the intervention of C (0.2MPa group, E group, 1H booster recovery (0.3MPa) turbocharged 1H recovery) increases the activity of skeletal muscle of rats with LDH; group G (0.2MPa turbocharged 1H recovery after hypoxic exercise group (F) and natural recovery after hypoxic exercise) significantly compared (P0.05) shows that the increases. Pressure assisted method intervention recovery group upregulation of LDH activity can promote skeletal muscle lactate to pyruvate metabolism, this process can reduce the content of lactic acid in skeletal muscle, increase of muscle energy supply, helps the body fatigue recovery in the plateau hypoxia environment after exercise.
(4) there was no significant change in SDH activity in skeletal muscle of rats after booster assisted intervention, indicating that booster assisted method after high altitude hypoxia had little effect on improving aerobic metabolism ability in rats.

【学位授予单位】：青海师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：G804.7

【参考文献】