【作者】 石真玉；

【导师】 肖国强；

【作者基本信息】 华南师范大学 ， 运动人体科学， 2004， 硕士

【摘要】 热休克蛋白或应激蛋白能保护细胞免受应激损伤，修复和维持蛋白的折叠，提高细胞在恶劣环境下的生存率。热应激能使机体产生热耐受性，同时合成大量的HSP70。热耐受性会对运动应激产生耐力，从而使机体在运动应激中得到保护。运动过程中自由基的爆发是导致机体损伤的原因之一。许多实验已经表明运动可以诱导骨骼肌和其他组织中HSP70的表达，但是不同的运动方式使得实验结果不尽相同。 本实验的目的是通过观察一次递增负荷的力竭运动对白细胞HSP70表达的影响，观察运动后恢复期HSP70的变化；给受试者热预处理，即发生热休克反应，观察HSP70在运动后的变化，同时检测机体的抗氧化能力及损伤情况，从而探讨在递增负荷运动中，热预处理诱导的HSP70高表达对机体的保护作用。 10名受试者在跑台上采用Bruce方法进行递增负荷运动至力竭。安静、运动后即刻(即3min)，运动后3h，运动后8h分别采肘静脉血，测试白细胞HSP70，血浆超氧化物歧化酶(SOD)、丙二醛(MDA)，另外测运动后即刻的血乳酸及血浆CK。间隔一周，热预处理1h(桑拿，温度45℃，湿度90％)，室温(温度25℃，湿度65％)恢复24h，采安静时肘静脉血测试。进行递增负荷运动至力竭后，采血测试各项指标。运动方式及采血时间同上。 实验结果： (1)运动后即刻白细胞HSP70与基础值比较没有显著性差异，运动后3h白细胞HSP70表达显著增加(P＜0.05)，8h已基本恢复到基础值。热预处理24h后HSP70显著增加，运动后即刻HSP70与热预处理前运动后即刻相比显著降低，8h的HSP70比基础值显著降低(P＜0.05)。华南师范大学体育科学学院2004届硕十学位论文石真玉 (2)两次运动后即刻血浆SOD活性都显著上升(P<0.05)，运动后3h与基础值比较无显著性差异，热预处理前各时刻SOD和热预处理后各时刻相比没有明显变化。运动后即刻血浆MDA含量显著上升，3h恢复到基础水平。热预处理运动后即刻MDA含量也明显上升，但是明显低于热预处理前运动后即刻(P<0.05)。 (3)运动后即刻血浆CK活性显著升高。热预处理运动后即刻CK活性没有明显变化，但显著低于热预处理前运动后即刻(P<0.05)。 (4)血乳酸，最大吸氧量，运动时间都没有显著性的变化。实验结果表明: (1)一次递增负荷的力竭运动能诱导机体白细胞HSP70的表达，且表达有延后J吐，既运动刺激结束后过一定的时间HSP70才表达增多;HSP70表达恢复至基础水平较快。热预处理运动后8h白细胞HSP70比基础值显著降低，提示HSP7O可能发生了适应性的变化。 (2)一次递增负荷的力竭运动可引起血浆SOD活性的升高，MDA含量增多，说明该运动方案虽然可能使机体抗氧化能力有所提高，但不能有效清除自由基，提示可能发生氧化损伤。(3)一次递增负荷的力竭运动可引起血浆中CK活性的提高，说明机体可能发生了一定的组织损伤。(4)对受试者施以热预处理而造成HSP7O高表达后，发现力竭运动后即刻MDA含量比热预处理前运动后即刻的MDA降低，SOD的活性没有明显的变化，提示热预处理诱导HSP70高表达可能对减少运动中自由基的产生有一定的作用。(5)热预处理运动后的CK活性比热预处理前运动的CK活性明显降低，说明热预处理后再运动对机体造成的损伤比热预处理前运动小，提示热预处理造成的HSP7O的高表达可能对机体的抗损伤有一定的作用。更多还原

【Abstract】 Heat shock proteins or stress proteins are considered to play an essential role in protecting cells from stress,repairing and maintaining protein folding and preparing them to survive new environmental challenges.HSP70 overexpression induced by heat stress is able to provide thermotolerance.This induces cellular adaptations which enable the organism to better deal with any future exercise challenge.Previous studies have demonstrated that a single exercise bout is capable of increasing HSP70 content.The purpose of this study was to observe HSP70 change after an incremental exhaustive bout of exercise,and HSP70 change postexercise after previous hyperthermic treatment.At the same time we examined antioxidant capacity and damage of organism and analyzed protection of HSP70 overexpression induced by hyperthermic pretreatment.10 male subjects of physical education department ran on a treadmill until exhaustion .Blood was sampled from forearm vein at rest,immediately postexercise,3h and 8h after exercise.These samples were analyzed for leucocytes HSP70 with Western dot blotting.Malonaldehyde (MDA) as a marker of lipid peroxidantion and plasma superoxide dismutase (SOD) were measured at rest,immediately postexercise and 3h after exercise. In addition,plasma creatine kinase(CK)was measured at rest and immediately postexercise as a crude marker of tissue damage.After a week the hyperthermic pretreatment was performed for an hour ( Sauna,45 癈 ;relative humidity,90%) .And then the subjects were at rest under the normal temperature for 24 hours.The same bout of exercise took place after this.The results were as follows: ( 1 ) HSP70 content in human leucocytes remained unchanged immediatelypostexercise and increased respectively from rest at 3h after exercise. It indicated that a single short-term exhaustive bout of exercise was able to increase leucocyte HSP70 expression. And it returned to pre-exercise level until 8h after exercise. HSP70 expression increased significantly at 24h after hyperthermic pretreatment.Compared with the level immediately postexercise before hyperthermic treatment,HSP70 decreased significantly immediately postexercise.HSP70 content at 8h after hyperthermic pretreatment decreased respectively compared with the basal line.This demonstrated that probably the adaptive response of the basal HSP expression took place.(2) SOD activity in plasma elevated significantly immediately after exercise.There were no significant differences between before hyperthermic treatment and after hyerthermic pretreatment.It is concluded that HSP70 overexpression induced by hyperthermic treatment is not able to enhance SOD activity in plasma.(3) Plasma MDA content increased respectively immediately postexercise.MDA elevated significantly immediately postexercise after hyperthermic pretreatment and it was lower respectively than that immediately postexercise before hyperthermic treatment.We conclude that HSP70 is probably involved in eliminating free radicals .(4) There was significant change of CK activity in plasma immediately after exercise. Plasma CK activity remained unchanged immediately postexercise after hyperthermic pretreatment,but was lower respectively than that immediately postexercise before hyperthermic treatment.This result demonstrates that HSP70 overexpression induced by hyperthermic treatment probably enables the organism to enhance the capacity of resistance to damage induced by exercise .(5) There were no significant differences in blood lactate,VO2max and exercise time.This indicates that HSP70 is not able to increase tolerance to a single short-term exhaustive exercise.更多还原