高压氧对递增负荷训练大鼠腓肠肌P53、Bcl-2蛋白表达的影响

背景：研究显示高压氧疗法在运动损伤与疲劳恢复方面具有较好的应用效果，但其机制尚不清楚。 目的：观察高压氧对递增负荷跑台运动后大鼠腓肠肌P53、Bcl-2蛋白表达及细胞凋亡的影响。 设计、时间及地点：随机对照动物实验，于2008-03/05在苏州大学运动人体科学实验室完成。 材料：2个月龄雄性SD大鼠24只，随机分为3组，安静对照组、递增负荷训练组、高压氧恢复组，每组8只。 方法：递增负荷训练组：坡度10%，递增负荷训练，第1~4周，每天20 m/min运动10 min，25 m/min运动10 min，30 m/min运动10 min，35 m/min运动10 min。第5~8周，每天20 m/min运动10 min，25 m/min运动10 min，30 m/min运动20 min，35 m/min运动20 min。高压氧恢复组：运动方案相同，运动后即刻用0.2 MPa压力的高压氧恢复60 min。安静对照组自由喂养，不进行训练。递增负荷训练组末次运动后1 h、高压氧舱恢复组恢复1 h及相应安静对照组分批麻醉处死，快速取腓肠肌立即用体积分数为0.1甲醛固定，制作切片。 主要观察指标：①苏木精-伊红染色观察各组大鼠骨骼肌细胞凋亡情况。②免疫组织化学检测各组大鼠骨骼肌细胞P53与Bcl-2蛋白表达情况。 结果：24只大鼠进入结果分析。①递增负荷训练组大鼠骨骼肌细胞凋亡数量增多，经高压氧恢复后细胞凋亡数量减少。②递增负荷训练组大鼠骨骼肌细胞P53蛋白表达增多，经高压氧恢复后表达显著减少。③递增负荷训练组大鼠骨骼肌细胞Bcl-2蛋白表达增多，高压氧恢复后未见减少。 结论：高压氧能够抑制递增负荷训练大鼠骨骼肌P53蛋白的表达，减少运动导致的大鼠骨骼肌细胞凋亡的发生。

Hyperbaric oxygen effects on the expression of P53 and Bcl-2 protein of gastrocnemius muscle in rats following incremental training

BACKGROUND: Hyperbaric oxygen therapy achieves excellent results in treating with sports injury and fatigue recovery, but its mechanism remains unclear. OBJECTIVE: To observe the influence of the expression of P53 and Bcl-2 protein after incremental training in gastrocnemius muscle and apoptosis of rats with hyperbaric oxygen. DESIGN, TIME AND SETTING: The ramdomized control animal experiment was performed in the Laboratory of Sports and Human Science, Suzhou University between March and May in 2008. MATERIALS: Twenty-four male Sprague-Dawley rats were randomly divided into three groups: control group, incremental training group, and hyperbaric oxygen group with 8 rats in each group. METHODS: Rats in the incremental training group were incremental training at 10% grade gradient with the speed of 20 m/min, 25 m/min, 30 m/min, 35 m/min, each speed exercise lasted for 10 min from 1 to 4 weeks. Then the duration of exercise incresed to 20 min at the 30m/min, and 35 m/min and 10 min in 20 m/min, 25 m/min in the next 4 weeks. Rats in the hyperbaric oxygen group rest in hyperbaric oxygen with the pressure of 0.2 MPa for 60 min after the training at the same motion scheme. Rats in the control group were feeding freely without training. Each group anesthetized rats were killed after exercise/rest 1 hour, and quickly picked out the gastrocnemius muscle, fasten with the 100 g/L formaldehyde, then made to slices. MAIN OUTCOME MEASURES: The gastrocnemius muscle apoptosis was observed by hematoxylin-eosin staining, expression of p53 and Bcl-12 were detected by immunohistochemistry. RESULTS: Totally 24 rats entered the final analysis. The number of gastrocnemius muscle apoptosis and P53 protein expression increased in the incremental training group, and decreased after recovery by the hyperbaric oxygen group respectively. Expression of Bcl-2 protein increased in the incremental training group, and without change in the hyperbaric oxygen group. CONCLUSION: Applying hyperbaric oxygen can inhibit the expression of P53 protein, and decrease the cell apoptosis of skeletal muscle caused by sport damage.