The purpose of this study was to determine the changes of both oxidant and antioxidant levels with exercise training in obese middle-aged women. The association between telomere length and oxidative stress with exercise was also examined. Sixteen obese middle-aged women participated in this study. The subjects were randomly divided into exercise group (EX) and control group (CON). EX performed aerobic exercise training for 6 months. DNA was extracted from leukocytes in peripheral blood and their telomere lengths were measured by real time PCR analysis. Long-term exercise training decreased body weight and BMI, and increased VO2 max. Resting levels of erythrocyte glutathione peroxidase activity were higher in EX compared to CON. Superoxide dismutase (SOD) activities were higher after the acute exercise test at mid-intensity in post-exercise training than in the pre-exercise training conditions. The telomere length did not change significantly after the acute exercise test in the pre-exercise training condition in spite of the increased level of malondialdehyde (MDA) as a marker of oxidative stress. In conclusion, antioxidant enzyme activities were increased following long-term exercise training; however, the lengths of telomere in leukocytes were not influenced by both mid-intensity and high intensity of exercise stress.