Background: The present study aimed to explore the effect of melatonin administration on lipid peroxidation in various tissues of rats with streptozocin induced diabetes and subjected to an acute swimming exercise. Methods: The study used 80 adult male rats, which were equally allocated to 8 groups: Group 1, general control; Group 2, melatonin-administered control; Group 3, melatonin-administered diabetic control; Group 4, swimming control; Group 5, melatonin-administered swimming; Group 6, melatonin-administered diabetic swimming; Group 7, diabetic swimming; Group 8, diabetic control. Diabetic rats were administered 3 mg/kg/day ip melatonin for 4 weeks. At the end of the study, the animals were decapitated to collect samples from liver, lung and spleen tissues, which were then analyzed to determine levels of liver MDA (nmol/gram/protein) and GSH (mg/g/protein). Results: The highest MDA values in liver, lung and spleen tissues were obtained in the Group 7. The values in the Group 8 were lower than those in the Group 7, but higher than in all other groups. The Group 5 and 6 had the highest liver, lung and spleen GSH values. Conclusion: Results obtained from the study indicate that the increase in free radical production and the inhibition of antioxidant activity in diabetes and acute exercise are both prevented by melatonin administration.