Visfatin produced by adipose tissue, skeletal muscle, liver and has insulin-mimetic actions. The studies shows, regular exercise enhances insulin sensitivity. Because the physiologic role of visfatin in exhaustive aerobic exercise responses are still unclear; The purpose of this study the effect of exhaustive aerobic exercise on responses of serum visfatin and insulin resistance in overweight middle-aged men trained.Eight healthy trained (age, 47.03±6.69 years; Weight 78.92±6.93 kg; height 170±7.34 cm; Vo2max 42.17±7.30 ml.kg-1.min-1 and body mass index 26.05±1.30 kg/m2); volunteers participated in the current study. Thus, subjects completed an informed consent form and health history questionnaire.The Exercise protocol included was Bulk test. Venous blood samples were collected before, immediately after exercise to assess serum visfatin, fasting glucose and insulin resistance responses. Data were analyzed by the Paired t-test and independent t- test.The results show that serum visfatin significant increased after exhaustive exercise (Pre 8.61±0.61 vs Post 10.51±1.57; P=0.031); while it remained unchanged in insulin resistance (Pre 1.19±0.36 vs Post 1.10±0.41; P=0.58). Therefore, promotion in physical activity level could be an effective way to increase visfatin and decrease overweight in middle-aged men trained.