Abstract Objective: In today’s world, one of the waves that humans have to deal with today, 2.45-GHz radio-frequency radiation (RFR) of wireless devices connected to the Internet, which can have various negative effects on human health. This study investigated sperm motility and male sex hormones NMRI Mice during long-term exposure to Wi-Fi radiation. Materials and Methods: Two-month-old NMRI male mice (30± 2 g) (n=27) were exposed to 2.45 GHz radiation in a room with two Wi-Fi antennas on the walls. Animals have been divided into 3 groups: I: check group (n=9), including healthy animals not exposed to the antenna, II: 4-hour group (n=9) exposed to the 2.45 GHz Wi-Fi radiation for 4 hour per day during two months and III: 8-hour group (n=9) exposed to the 2.45 GHz Wi-Fi radiation for 8 hours per day during 3 months. Sperm motility and sex hormone levels were evaluated in both exposed and control animals. Results: Sperm motility and average hormone concentration Testosterone and LH levels decreased substantially in the 8-hour groups compared to the control group. However, these parameters were not significantly reduced in the group (4 hours/day/2 months) compared to the control group.