运动对大鼠骨骼肌FOS和HSP70表达的影响研究
摘要
目的:通过实验观察一次急性运动和长期耐力运动前、中、后不同时期大鼠骨骼肌FOS蛋白和HSP70表达情况,找出急性和慢性运动两种运动情况下FOS蛋白和HSP70表达规律从而推断二者之间是否具有相关性。
方法:选取健康雄性SD大鼠100只,随机分成两大组:一次急性运动组和长期耐力组。参考Bedford渐增负荷跑台运动模型和常芸的长期耐力跑台模型进行造模,造模结束后按照实验设计对应时间点进行骨骼肌取材,常规脱水、石蜡包埋、HE染色。FOS蛋白和HSP70采用免疫组织化学法进行染色。用LEICA Qwin图像分析系统测定FOS蛋白和HSP70阳性表达面积百分比。
结果:(1)一次急性运动后大鼠血清CK活性在运动后5h显著升高(P＜0.05),运动后12h达到高峰,并且一直持续到运动后24h(P＜0.05)。在长期耐力组中,安静对照组、运动后即刻组、运动后24h三者之间均无显著性差异。
(2)FOS一次急性运动组中,安静对照组几乎没有阳性表达,运动后即刻阳性表达显著增多(P＜0.01),1h达到高峰(P＜0.01),一直持续到运动后3h。运动后5h渐下降,运动后24h显著高于安静对照组(P＜0.05)。在长期耐力组中,运动后即刻组与安静对照组有显著性(P＜0.05),运动后24h恢复至安静水平。
(3)HSP70一次急性运动组中,安静对照组有少量阳性表达,运动后即刻阳性表达显著增多(P＜0.05),运动后1h阳性面积进一步增大(P＜0.01),运动后3h达到高峰,一直持续到运动后5h(P＜0.01)运动后24h始下降,但仍显著高于安静对照组(P＜0.05)。在长期耐力组中,运动后即刻阳性表达显著增多(P＜0.01),运动后24h低于运动后即刻但与安静对照组相比有显著性(P＜0.01)。
结论:本研究得出以下结论(1)一次急性运动造成了大鼠腓肠肌肌纤维排列紊乱,肌细胞轻度充血水肿;而长期耐力训练后大鼠腓肠肌纤维明显增粗,细胞间质减少,结构致密。(2)一次急性运动可以引起大鼠血清CK在24h内发生显著变化。但长期耐力运动后血清CK在24h内没有明显变化。(3)运动可以显著诱导c-fos及HSP70表达,FOS蛋白及HSP70表达量的增加与运动损伤有关。(4)c-fos基因在应激后能迅速表达,1h达到高峰。HSP70表达迟于FOS蛋白,峰值在运动后3h到来,FOS蛋白表达可能诱导了HSP70的表达。
Objective: To observe FOS and heat shock protein expression in rat skeletal muscle before, during and after different periods of acute and long-term endurance exercise, we try to identify in two different exercise circumstances whether FOS and heat shock protein have relativity or not.
Methods: 100 male SD rats were randomly divided into two groups: acute and long-term endurance exercise group. As the incremental load running with Bedford treadmill running model and treadmill runing model of YunChang referenced, rats were killed accordance with points of design in experiment after last exercise. The change of skeletal muscle was viewed by HE dyeing. The expression of FOS and HSP70 were detected by immunohistochemistry stain. Using LEICA Qwin pictures analyze system measured the area of positive cell of FOS and HSP70 expression. The data were analyzed by SPSS 13.0.
Results: (1) Activities of CK in serum in skeletal muscles of rats after acute exercise were significantly higher (P <0.05) after five hours, the movement reached a peak at 12 hours after exercise, and continued until 24 hours after exercise (P <0.05) . In the long-term endurance group, after 10 weeks of training three group have no significant difference.2) in the acute group, the rest control group have no positive cells, but in immediately after exercise group positive cells significantly increased (P <0.01), one hour after exercise the peak appeared (P <0.01), and continued until three hours after exercise, After five hours it has been gradually declining but still significantly higher than the quiet control group 24 hours after exercise (P <0.05). In the long-term endurance group, the group immediately after exercise positive cells and quiet control rest group was significant (P <0.05) after 24 hours positive movement returned to the level of quiet levels.(3) HSP70 in the acute exercise group, the control rest group, there are quiet a small number of positive cells, the movement of immediately after exercise group positive cells significantly increased (P <0.05), one hour after exercise group was further increased (P <0.01), the exercise of the 3 reached a peak hour, has continued to campaign after 5 hours, 24 hours after the campaign started to decline, but still significantly higher than the quiet control group (P <0.05). In the long-term endurance group, the Movement immediately after exercise group positive cells keep significantly increased (P <0.01) Until 24 hours after exercise.
Conclusion: In this study we have come to conclusions (1) an acute movement caused rat gastrocnemius muscle fiber with disorder, muscle cells, such as Swelling and Congestive, while long-term endurance training rat gastrocnemius fiber is obviously enlarged, stromal cells reduce and the structure is more dense.(2)an acute movement can cause serum CK significant changes in rats at 24 h after exercise. But long-term endurance exercise after 24 h of serum CK has not changed. (3)Exercise can significantly induced c-fos and HSP70 expression in skeletal muscle of rats . sports injury can increased expression of HSP70 and c-fos. (4) FOS expression quickly after exercise it can reached a peak at 1 hour after exercise. HSP70 can reached a peak at 3 hour after exercise. FOS protein maybe induces the expression of HSP70 expression.
方法:选取健康雄性SD大鼠100只,随机分成两大组:一次急性运动组和长期耐力组。参考Bedford渐增负荷跑台运动模型和常芸的长期耐力跑台模型进行造模,造模结束后按照实验设计对应时间点进行骨骼肌取材,常规脱水、石蜡包埋、HE染色。FOS蛋白和HSP70采用免疫组织化学法进行染色。用LEICA Qwin图像分析系统测定FOS蛋白和HSP70阳性表达面积百分比。
结果:(1)一次急性运动后大鼠血清CK活性在运动后5h显著升高(P＜0.05),运动后12h达到高峰,并且一直持续到运动后24h(P＜0.05)。在长期耐力组中,安静对照组、运动后即刻组、运动后24h三者之间均无显著性差异。
(2)FOS一次急性运动组中,安静对照组几乎没有阳性表达,运动后即刻阳性表达显著增多(P＜0.01),1h达到高峰(P＜0.01),一直持续到运动后3h。运动后5h渐下降,运动后24h显著高于安静对照组(P＜0.05)。在长期耐力组中,运动后即刻组与安静对照组有显著性(P＜0.05),运动后24h恢复至安静水平。
(3)HSP70一次急性运动组中,安静对照组有少量阳性表达,运动后即刻阳性表达显著增多(P＜0.05),运动后1h阳性面积进一步增大(P＜0.01),运动后3h达到高峰,一直持续到运动后5h(P＜0.01)运动后24h始下降,但仍显著高于安静对照组(P＜0.05)。在长期耐力组中,运动后即刻阳性表达显著增多(P＜0.01),运动后24h低于运动后即刻但与安静对照组相比有显著性(P＜0.01)。
结论:本研究得出以下结论(1)一次急性运动造成了大鼠腓肠肌肌纤维排列紊乱,肌细胞轻度充血水肿;而长期耐力训练后大鼠腓肠肌纤维明显增粗,细胞间质减少,结构致密。(2)一次急性运动可以引起大鼠血清CK在24h内发生显著变化。但长期耐力运动后血清CK在24h内没有明显变化。(3)运动可以显著诱导c-fos及HSP70表达,FOS蛋白及HSP70表达量的增加与运动损伤有关。(4)c-fos基因在应激后能迅速表达,1h达到高峰。HSP70表达迟于FOS蛋白,峰值在运动后3h到来,FOS蛋白表达可能诱导了HSP70的表达。
Objective: To observe FOS and heat shock protein expression in rat skeletal muscle before, during and after different periods of acute and long-term endurance exercise, we try to identify in two different exercise circumstances whether FOS and heat shock protein have relativity or not.
Methods: 100 male SD rats were randomly divided into two groups: acute and long-term endurance exercise group. As the incremental load running with Bedford treadmill running model and treadmill runing model of YunChang referenced, rats were killed accordance with points of design in experiment after last exercise. The change of skeletal muscle was viewed by HE dyeing. The expression of FOS and HSP70 were detected by immunohistochemistry stain. Using LEICA Qwin pictures analyze system measured the area of positive cell of FOS and HSP70 expression. The data were analyzed by SPSS 13.0.
Results: (1) Activities of CK in serum in skeletal muscles of rats after acute exercise were significantly higher (P <0.05) after five hours, the movement reached a peak at 12 hours after exercise, and continued until 24 hours after exercise (P <0.05) . In the long-term endurance group, after 10 weeks of training three group have no significant difference.2) in the acute group, the rest control group have no positive cells, but in immediately after exercise group positive cells significantly increased (P <0.01), one hour after exercise the peak appeared (P <0.01), and continued until three hours after exercise, After five hours it has been gradually declining but still significantly higher than the quiet control group 24 hours after exercise (P <0.05). In the long-term endurance group, the group immediately after exercise positive cells and quiet control rest group was significant (P <0.05) after 24 hours positive movement returned to the level of quiet levels.(3) HSP70 in the acute exercise group, the control rest group, there are quiet a small number of positive cells, the movement of immediately after exercise group positive cells significantly increased (P <0.05), one hour after exercise group was further increased (P <0.01), the exercise of the 3 reached a peak hour, has continued to campaign after 5 hours, 24 hours after the campaign started to decline, but still significantly higher than the quiet control group (P <0.05). In the long-term endurance group, the Movement immediately after exercise group positive cells keep significantly increased (P <0.01) Until 24 hours after exercise.
Conclusion: In this study we have come to conclusions (1) an acute movement caused rat gastrocnemius muscle fiber with disorder, muscle cells, such as Swelling and Congestive, while long-term endurance training rat gastrocnemius fiber is obviously enlarged, stromal cells reduce and the structure is more dense.(2)an acute movement can cause serum CK significant changes in rats at 24 h after exercise. But long-term endurance exercise after 24 h of serum CK has not changed. (3)Exercise can significantly induced c-fos and HSP70 expression in skeletal muscle of rats . sports injury can increased expression of HSP70 and c-fos. (4) FOS expression quickly after exercise it can reached a peak at 1 hour after exercise. HSP70 can reached a peak at 3 hour after exercise. FOS protein maybe induces the expression of HSP70 expression.
引文
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