摘要
目的:观察过度训练及补充二联甲苯(DPI)或谷氨酰胺(Gln)对大鼠腹膜巨噬细胞活性氧(ROS)生成能力的影响,探讨诱导型一氧化氮合酶(iNOS)在此过程中发挥的作用。方法:8周龄健康雄性wistar大鼠56只,随机分为安静对照组(C)、过度训练组(E)、过度训练补充DPI组(ED)、过度训练补充Gln组(EG)。后3组根据取材时间不同各分为2组,运动后36h取材组(E1、ED1、EG1),运动后7天取材组(E2、ED2、EG2)。总计7组,每组8只,除C组外,其他6组都进行11周递增负荷跑台训练。断头处死大鼠并分离纯化腹膜巨噬细胞,流式细胞术测定ROS生成量,荧光定量PCR技术测定iNOS基因表达。结果:ROS生成:E1组显著低于C组;ED1组显著低于E1组(P<0.05),与C组相比极显著降低(P<0.01);EG1组与E1组相比没有显著差异,与C组相比极显著降低(P<0.01)。E2组、ED2组、EG2组分别与E1组、ED1组、EG1相比均显著增高;E2组、ED2组、EG2组、C组4组之间比较无显著差异。iNOSmRNA表达:ED1组相对E1组显著增加(P<0.05),相对C组极显著增加(P<0.01);ED2组相对ED1组显著降低;其他相关组间比较无显著差异。对各组中ROS与iNOS表达量进行相关分析,显示二者呈直线负相关,相关系数r=-0.45(P=0.004)。以各组ROS和iNOS表达量相对,C组的倍比关系作图,发现ROS生成量较高时,iNOSmRNA表达处于较低水平(E2组),随着iNOSmRNA表达水平增加,ROS生成量呈现降低趋势。结论:过度训练使腹膜巨噬细胞ROS生成量显著低于生理水平,引起运动性免疫抑制;补充抗氧化剂DPI可使过度训练时巨噬细胞ROS生成量进一步降低,补充谷氨酰胺对过度训练引起的巨噬细胞ROS生成量降低没有改善作用;过度训练及补充DPI或谷氨酰胺可影响巨噬细胞内ROS生成,这在一定程度上受iNOS催化产生的NO调控。
Objective：To investigate the effect of overtraining and supplement with DPI（diphenylene iodonium） or glutamine on the production of ROS（reactive oxygen species） in pritoneal macrophages and the function of iNOS（inducible nitric oxide synthase） in this process.Method：56 male wistar rats were randomly divided into 7 groups：sedentary group（C,n=8）,overtraining group（E）,overtraining supplement with DPI group（ED）,overtraining supplement with Gln group（EG）.E,ED and EG group were respectively divided into two groups which sacrificed at 36h（E1,ED1,EG1,n=8） and 7 days（E2,ED2,EG2,n=8） after the last training.All groups except C were training in standard treadmill with an increasing load for 11 weeks.Peritoneal macrophages were isolated and purified after all rats were sacrificed by decapitation.The production of ROS was detected by FACS,and real time-PCR was used to detect the mRNA expression of iNOS.Result：Overtraining group showed a significant decline in production of ROS when compared with pritoneal macrophage from the sedentary group,and ED1 group significant lower than E1 group（P0.05） and sedentary group（P0.01）.The production of ROS in EG1 group had no significant difference when compared with overtraining group,however,it significant lower than the sedentary group（P0.01）.E2,ED2 and EG2 group showed a significant increase when compared with E1,ED1 and EG1 group respectively.There were no significant differences in E2,ED2,EG2 and C group.The expression of iNOS in ED1 group significant higher than overtraining group（P0.05）,sedentary group（P0.01） and ED2 group（P0.05）.The iNOS mRNA levels had no significant differencies in E2,ED2,EG2 and C group.The correlation analysis showed that ROS and iNOS were negative correlation and the pearson correlation was-0.45（P=0.004）.When the production of ROS was highness,the expression of iNOS was the lowest（E2 group）.When the iNOS mRNA levels increase,the production of ROS showed a tendency of decline.Conclusion：Overtraining made the production of ROS in pritoneal macrophage lower than physiological level significantly,which was the performance of exercise-induced immunosuppression.DPI supplement made the production of ROS in overtraining group decline more seriously.And the decline of ROS in overtraining group could not be ameliorated by Gln supplement.Overtraining and supplement with DPI or Gln certainly had an influence on ROS production in pritoneal macrophages,and it was regulated by NO which mediate by iNOS in a certain extent.
出处
《体育科学》
CSSCI
北大核心
2011年第2期49-54,共6页
China Sport Science
基金
国家自然科学基金项目(30971422)
上海市重点学科建设资助项目(S30802)
