We investigated the effects of melatonin administration on skeletal muscle ischaemia-reperfusion injury (IRI) by assessing plasma malondialdehyde (MDA), superoxide dismutase (SOD), total glutathione (GSSH), glutathione peroxidase (GPX) and myeloperoxidase (MPO) concentrations. Male Sprague-Dawley rats (n = 32) were randomized into four groups: group 1 served as time controls; group 2 were the test animals; group 3 received melatonin (30 mg/kg) intraperitoneally prior to the induction of ischaemia; and group 4 received melatonin (30 mg/kg) intraperitoneally prior to the reperfusion period. Administration of melatonin prior to reperfusion significantly decreased the elevated MDA concentration caused by IRI, and significantly elevated GSSH concentrations, which had been reduced by IRI. Ischaemia-reperfusion injury significantly increased activities of GPX, SOD and MPO, and melatonin administration reversed this effect. In conclusion, a pharmacological dose of melatonin showed significant protective effects against IRI by decreasing lipid peroxidation, MPO, SOD and GPX enzyme activities and regulating glutathione content.