Effects of Overtraining and Supplement with DPI or Glutamine on the Production of ROS and the Expression of iNOS in Pritoneal Macrophages

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(P<0.05) and sedentary group(P<0.01).The production of ROS in EG1 group had no significant difference when compared with overtraining group,however,it significant lower than the sedentary group(P<0.01).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(P<0.05),sedentary group(P<0.01) and ED2 group(P<0.05).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.