摘要
目的:利用电刺激使C2C12细胞产生收缩运动,实时观察细胞内活性氧(ROS)生成的时相变化。方法:利用培养的C2C12细胞,电刺激使其产生收缩运动,测定细胞ROS生成速率、线粒体MDA含量、总SOD活性和胞浆内Na+,K+-ATPase活性。结果:(1)以45V、20ms5、Hz的强度刺激C2C12细胞,细胞内ROS生成迅速增加,在刺激60min时ROS的生成速率呈显著性升高(P<0.01),然后缓慢下降,继续刺激到120min时,ROS生成再次升高(P<0.01),随后迅速下降。(2)线粒体总SOD活性逐渐升高,至150min和180min时都与对照组相比有显著性差异(P<0.05);MDA量在90min时略有升高,随后下降,但无显著性差异。(3)电刺激C2C12细胞引起胞浆内Na+,K+-ATPase活性升高,刺激至180min时与刺激90min时相比酶活性显著下降(P<0.05)。结论:利用电刺激C2C12细胞模型,可实时测定收缩细胞的ROS生成速率变化,为运动性ROS产生的机理提供了直接证据。
Objective To investigate the time course of ROS generation and the change of antioxidant system in electrical - stimulated C2C12. Methods In this study, C2C12 was stimulated electrically in order to induce the contraction of the cell vitro. ROS was detected with DCFH fluorescent probe. Lipid peroxides （MDA）, superoxide dismutase （SOD） and Na^＋ , K^＋ -ATPase activities were also detected. Results （1） Rapid increase in ROS production during stimulation was found and the peak revealed 60min after stimulation （ P 〈 0.01 ）, than ROS production decreased rapidly. The ROS in C2C 12 cell increased once again as stimulation continued and the peak revealed 120min after stimulation（ P 〈 0.01）. （2）Mitochondrial SOD activity increase gradually and were significantly higher than baseline at 150min and 180min（ P 〈 0.05） after stimulation. Increase and decrease in MDA were also observed without statistical difference. （3）The activity of Na^＋ , K^＋ - ATPase was lower at 180min than at 90min after the stimulation（ P 〈 0.05）. Conclusions The model used in this study could be useful in the direct real - time measurement of ROS generation, and the results of the study provided the direct evidence on the mechanism of exercise - induced ROS generation.
出处
《中国运动医学杂志》
CAS
CSCD
北大核心
2006年第1期46-49,共4页
Chinese Journal of Sports Medicine
基金
广东省自然科学基金(31526)资助
