The aim of this study was to examine the effect of high-intensity sprint and strength training (HISST) on glucoregulatory hormones in young (20 years) and middle-aged (40 years) men. Thirty-six moderately trained men participated as volunteers in this study. After medical examination, eligible subjects were randomly assigned to one of four groups according to their age: a young training group (21.3 ± 1.3 yrs, YT, n = 9), a young control group (21.4 ± 1.7 yrs, YC, n = 9), a middle-aged training group (40.7 ± 1.8 yrs, AT, n = 9), and a middle-aged control group (40.5 ± 1.8 yrs, AC, n = 9). YT and AT participated in HISST for 13 weeks. Before and after HISST, all participants performed the Wingate Anaerobic Test (WAnT). Blood samples were collected at rest, after warm-up (50% VO2max), immediately post-WAnT, and 10 min post-WAnT. Before HISST, we observed significantly higher (P < 0.05) glucose concentrations in AT (5.86 ± 0.32 mmol.L-1) compared to YT (4.24 ± 0.79 mmol.L-1) at rest, and in response to WAnT (6.56 ± 0.63 mmol.L-1 vs. 5.33 ± 0.81 mmol.L-1). Cortisol levels were significantly higher (P < 0.05) in AT than in YT in response to WAnT (468 ± 99.50 ng.mL-1 vs. 382 ± 64.34 ng.mL-1). Catecholamine levels measured at rest and in response to WAnT rose in a similar fashion. After HISST, this "age effect" disappeared at rest and in response to exercise in the trained groups (YT and AT). Changes in hormone concentrations with intense training are due to adaptive changes in various tissues, especially in the skeletal muscle and liver in trained subjects. HISST may, at least in part, counteract the negative "age effect" on glucose metabolism.