This study investigates the protective effects of Vitamin E and Melatonin against gamma ray-induced damage in rat testes. A total of 40 Wistar rats were divided into four groups. The first three groups were exposed to 360 cGy gamma radiation using a Cobalt-60 apparatus. After radiation exposure, the first group received Vitamin E (100 mg/kg/day), the second group received Melatonin (100 mg/kg/day), and the third group received 0.9% saline solution. The fourth group served as the control group and received only saline without radiation. Five days later, the rats’ testes were removed and analyzed for Malondialdehyde (MDA) levels and histopathological damage. The Melatonin group showed no significant difference in MDA levels and tissue damage compared to the control group (p>0.05). However, in the Vitamin E and saline groups, both MDA levels and tissue damage were significantly increased (p<0.05). In conclusion, Melatonin demonstrated a strong protective effect by preventing free ra