The effects of 24 weeks of high-intensity interval training (HIIT) on markers of male reproductive function in infertile patients were studied. Infertile men (n = 441) were randomized to exercise (EX, n = 221) or non-exercise (NON-EX, n = 220) group. Patients in the EX group performed an interval training (1:1 work:rest ratio) 3 times per week at 75-95% of maximal oxygen consumption, for 24 weeks (VO2max). Markers of inflammation and oxidative stress in the seminal plasma, as well as semen parameters, sperm DNA fragmentation and rates of pregnancy, were measured at baseline, on weeks 12, 24; and 7 and 30 days thereafter during the recovery period. The intervention resulted in decreased seminal levels of proinflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) and markers of oxidative stress (ROS, MDA, and 8-isoprostane) (P < 0.05). The concentrations of seminal antioxidants were unaltered with HIIT intervention. These changes further coincide with promising developments in semen parameters, sperm DNA integrity and rates of pregnancy (P < 0.05). This may indicate that HIIT induced beneficial effects on markers of male reproductive function through decreased oxidative damage and proinflammatory status. Findings highlight the possibility that HIIT may be an effective intervention for male factor infertility and support the need for further human studies. Copyright © 2019 Elsevier Ltd. All rights reserved.