作　　者： ; ;

机构地区： 上海体育学院运动科学学院

出　　处： 《体育科学》 2011年第1期29-38,共10页

摘　　要： 目的：探讨过度运动激活中性粒细胞NADPH氧化酶介导产生的活性氧（ROS）对淋巴细胞DNA损伤的影响及实施二联苯碘（DPI）干预作用。方法：30只雄性Vistar大鼠随机分为3组,安静组（C,n=10）、过度训练组（E,n=10）和DPI干预组（D,n=10）,E组和D组进行递增负荷跑台训练11周,末次训练结束后在36~40 h内各组随机取8只眼眶静脉采血。应用ELISA方法检测血浆细胞因子水平及血液中脂质过氧化水平;提取白细胞用An-nexinV/PI双染色法流式测定中性粒细胞凋亡与坏死程度;免疫细胞化学的激光共聚焦法测定NADPH氧化酶关键亚基的gp91 phox与p47 phox的共定位;单细胞凝胶电泳测定淋巴细胞的DNA损伤程度。结果：1）与C组比较,E组血浆IL-1 b显著性升高,IL-8、TNF-α显著升高（分别为P〈0.01和P〈0.05）,MCP-1与CINC显著性下降（P〈0.05）;而D组血浆IL-1 b、MCP-1显著下降（P〈0.05）,IL-8、TNF-α显著性升高（P〈0.05）;E组和D组大鼠血浆MDA、MPO水平均上升（分别为P〈0.01和P〈0.05）;2）与C组相比,E组、D组中性粒细胞性凋亡的百分比均显著上升（P〈0.01）,但中性粒细胞的死亡数E组增加（P〈0.05）而D组无显著变化;3）与C组比较,E组出现DNA损伤的彗星细胞数非常显著性升高（P〈0.01）,且彗星细胞的宽度和尾长的变化均达到显著性水平,而D组的变化均未达到显著性水平;4）中性粒细胞的共聚焦染色定位显示：与安静组对照比较,E组出现了NADPH氧化酶关键亚基的gp91 phox与p47 phox的共定位。结论：1）过度运动可引起外周血炎性因子和趋化分子分泌的增加,从而有可能诱发组织炎症的发生和而调节T淋巴细胞的分化;2）过度运动可激活外周血中性粒细胞的NADPH氧化酶介导产生过多的ROS使血液的脂质过氧化水平提高;3）由此途径产生的ROS的增多和血液过氧化水平的提高可能引起中性粒细胞本身的凋亡甚至死亡� Objective：To investigate the effects of reactive oxygen species（ROS） mediated by NADPH-oxidase in neutrophils and intervention of biphenyl iodide（DPI） on DNA damage of lymphocytes induced by over training.Methods：30 male vistar rats were randomly divided into three groups：blank control group（C,n=10）,overtraining group（E,n=10） and DPI intervention group（G,n =10）.Group E and G were training in standard treadmill with an increasing load for 11 weeks,within 36h-40h after the last training,eight rats were randomly pick out from each group and the orbital venous blood were obtained.ELISA was used to detect the level of cytokine and blood lipid peroxidation in blood plasma.Utilizing flow cytometer and AnnexinV / PI double staining to measure the apoptosis and necrosis of neutrophil,using immunocytochemistry and confocal microscopy to indentify the co-localization both gp91phox and p47phox,which is the key subunits of the NADPH-oxidase and single cell gel electrophoresis（SCGE） was used to detect DNA damage of lymphocytes.Results：1.Compared with group C,the concentration of IL-1b,IL-8,TNF-α were significantly increased and the MCP-1,CINC were significantly decreased in rat blood plasma of group E（respectively,P〈0.01 and P〈0.05）;the concentration of IL-1b,MCP-1 were decreased significantly（P〈0.05）,and IL-8,TNF-α were significantly increased（P〈0.05） in rat blood plasma of group D;the level of MDA,MPO were increased in plasma of group E and D（respectively P〈0.01 and P〈0.05）.2.Compared with group C,the percentage of apoptosis of neutrophils significantly increased（P〈0.01） in group E and D,and the percentage of cell death increased in groups E（P〈0.05） while no significant change in group D.3.Compared with group C,the number of comet cells,which indicate the DNA damage significantly increased（P〈0.01） and the width and the tail length of comet cells were significant increased in group E while no increase in group D.4.the co-location confocal of gp91p

关 键 词： 过度运动 中性粒细胞 活性氧 淋巴细胞 损伤 干预 鼠 动物实验

领　　域： [文化科学] [文化科学]