The current study examine the effects of combined sprint and resistance training on serum total testosterone (TT), sex-hormone-binding globulin (SHBG) and cortisol (C), at rest, and in response to the Wingate Anaerobic-Test (WAnT) in 21 and 41 years old men. Forty moderately-trained men were randomly assigned to a young trained (YT), young control (YC), middle-aged trained (MAT), and middle-aged control (MAC) group. Before (P1), and after (P2) training, blood samples were collected at rest and after exercise. At P1, higher C and lower TT was observed in middle-aged groups compared to younger ones (P < 0.05). At P2, basal TT increased significantly (P < 0.05) in MAT and the age-difference was absent between trained groups (P > 0.05). Basal SHBG decreased significantly in YT at P2 (P < 0.05) but did not change in other groups from before to after training (P > 0.05). Free-testosterone was significantly (P < 0.05) higher in young compared to middle-aged groups at P1, but at P2, this age-related difference disappeared between YT and MAT (P > 0.05). C post-WAnT increased significantly for MAT only (P < 0.05) at P2, whilst no significant changes were observed in the other three groups (P < 0.05) at P2. In contrast, resting levels of C did not change in all groups at P2 (P > 0.05). The current study demonstrates that this training intervention may help increase steroids hormones in middle-aged men and counteract the negative effect of age on TT and free testosterone.