急性递增负荷运动对小鼠骨骼肌SOD同工酶活性和mRNA表达的影响

doi:10.15942/j.jcsu.2016.03.021

成都体育学院学报 ›› 2016, Vol. 42 ›› Issue (3): 116-121.doi: 10.15942/j.jcsu.2016.03.021

急性递增负荷运动对小鼠骨骼肌SOD同工酶活性和mRNA表达的影响

王平¹, LIChunguang², 漆正堂^{3, 4}, 丁树哲⁴

1.杭州师范大学体育与健康学院,浙江杭州 311121;
2. National Institute of Complementary Medicine, University of Western Sydney, Penrith, New South Wales 2751, Australia;3.华东师范大学青少年健康评价与运动干预教育部重点实验室,上海 200241;4.华东师范大学体育与健康学院,上海 200241

收稿日期:2015-09-24 修回日期:2016-03-11 出版日期:2016-05-15 发布日期:2016-05-27
作者简介:王平,副教授,博士后,主要研究方向:骨骼肌运动适应与信号调控。E-mail:wppa7476@163.com。
基金资助:
国家自然科学基金资助项目“线粒体蛋白输入(PIM)的运动适应与调控机制研究”(31171142);浙江省教育厅资助项目“运动诱导自噬信号通路调控骨骼肌质量变化研究”(Y201328990);杭州师范大学博士启动金“运动诱导自噬信号通路调控骨骼肌质量变化研究”(PE13002004028)。

Effect of Acute Incremental Load Exercise on SOD activity and mRNA in Skeletal Muscle of Mice

WANG Ping¹, LI Chunguang², QI Zhengtang^3,4, DING Shuzhe⁴

1. School of Physical Education and Health,Hangzhou Normal University,Hangzhou Zhejiang 311121;
2. National Institute of Complementary Medicine, University of Western Sydney, Penrith, New South Wales 2751, Australia;
3.Key Laboratory of Adolescent Health Assessment and Exercise Intervention Ministry of Education,East China Normal University,Shanghai 200241;
4.School of Physical Education and Health,East China Normal University, Shanghai 200241

Received:2015-09-24 Revised:2016-03-11 Online:2016-05-15 Published:2016-05-27

摘要/Abstract

摘要： 目的:通过建立急性递增负荷小鼠运动模型,探讨小鼠急性跑台运动后骨骼肌抗氧化防御系统的反应。方法:30只6周龄清洁级雄性ICR小鼠,体重20.81±2.31g,随机分为安静对照组(C组,6只)、急性递增负荷运动45min组(E1组,6只)、90min组(E2,6只)、120min组(E3组,6只)和150min组(E4组,6只)组。正式运动按以下程序:第1 级负荷:0°,8.2m/min(相当于53%VO₂max),15min;第2 级负荷:5°,15m/min(相当于64%VO₂max),15min;第3 级负荷:10°,19.3m/min(相当于76%VO₂max)。各组分别运动至所设相应时间及安静对照组,取出左、右侧腓肠肌,采用硝酸还原酶法测定线粒体NO含量,采用化学比色法测定eNOS活性,采用黄嘌呤氧化酶法测定线粒体MnSOD、CuZnSOD的活性;实时荧光定量PCR(Real-time PCR)检测骨骼肌MnSOD、CuZnSOD基因表达水平。结果:(1)与安静对照组相比,小鼠骨骼肌线粒体NO含量在急性递增负荷运动45min、90min、120min和150min均出现显著增加(P<0.05),且90min达到峰值(P<0.01);小鼠骨骼肌线粒体eNOS活性在急性递增负荷运动45min、90min和120min均出现显著增加(P<0.05),且90min达到峰值(P<0.01),但150min未出现显著性变化(P>0.05);(2)线粒体MnSOD活性在急性运动45min后明显增加(P<0.05),急性运动90、120min和150min均出现极显著增加(P<0.01),且在120min达到峰值;(3)与安静对照组相比,MnSODmRNA在急性运动45min出现显著性增加(P<0.05),急性运动90min、120min和150min均出现极显著性增加(P<0.01),且在150min达到峰值(P<0.01)。结论:线粒体MnSOD对急性递增负荷运动具有较好的应答反应,是运动刺激-反应-适应的主要SOD机制。

关键词: 急性递增运动, 骨骼肌, NO, eNOS, MnSOD, CuZnSOD

Abstract: Objective: To investigate the changes of the content of NO and activity of eNOS,the mRNA expression and the activity changes of MnSOD and CuZnSOD on acute incremental exercisein skeletal muscle of mice and explore the effects of antioxidant defense system. Methods: thirty male ICR mice were randomly divided into control group (C), 45min exercise group (E1, n=6), 90min exercise group (E2, n=6), 120min exercise group (E3, n=6) and 150min exercise group (E4, n= 6). Mice in exercise groups were trained through treadmill exercise (0°grade, 5m/min, 5min/day) for 3 days. After adaptation, mice in exercise groups underwent treadmill exercise to the corresponding time set up. Exercise patterns are as follows: First, 8.2m/min, 0 grade, 15min; second, 15m/min, 5% grade, 15min; third, 19.3m/min,10% grade. At the end of exercise, all mice were killed and their gastrocnemius muscle were obtained. Left one was used as mitochondria extraction, right one was measured by Real-time PCR. Results: (1) Compared with control group,NO content significantly increased after 45min exercise (P<0.05), 90min((P<0.01)), 120min and 150min (P<0.05) and reached the peak at 90 min. eNOS activity significantly increased after 45min exercise (P<0.05), 90min(P<0.01), 120min (P<0.05) and reached the peak at 90min.But the eNOS activity didn’t increase in 150min group significantly (P<0.05);(2) MnSOD activity was significantly increased after 45min exercise (P<0.05), 90min, 120min and 150min (P<0.01) and reached the peak at 120min.But the CuZnSOD activity didn’t increase in all exercise groups significantly (P<0.05); (3) Compared with control group, MnSOD mRNA significantly increased after 45min exercise (P<0.05), 90min, 120min and 150min (P<0.01) and reached the peak at 150min.But the CuZnSOD mRNA didn’t increase in all exercise groups significantly (P<0.05).Conclusion: MnSOD is the key SOD enzyme in response to the acute incremental exercise, which forms a protective cycling mechanism in acuteskeletal muscle exercise.

Key words: Acute Incremental Exercise, Skeletal Muscle, NO, eNOS, MnSOD, CuZnSOD

中图分类号:

G804.7

王平, LIChunguang, 漆正堂, 丁树哲. 急性递增负荷运动对小鼠骨骼肌SOD同工酶活性和mRNA表达的影响[J]. 成都体育学院学报, 2016, 42(3): 116-121.

WANG Ping, LI Chunguang, QI Zhengtang, DING Shuzhe. Effect of Acute Incremental Load Exercise on SOD activity and mRNA in Skeletal Muscle of Mice[J]. Journal of Chengdu Sport University, 2016, 42(3): 116-121.

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急性递增负荷运动对小鼠骨骼肌SOD同工酶活性和mRNA表达的影响

Effect of Acute Incremental Load Exercise on SOD activity and mRNA in Skeletal Muscle of Mice

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