Abstract: The aim of this study was to evaluate the possible antioxidant effect of ghrelin against tilmicosin-induced myocardial damage in rats. Forty male Sprague Dawley rats were equally divided into four groups: control (saline for 5 days), tilmicosin (single dose of 75 mg kg‾1, s.c.), ghrelin (10 ng/kg/day for 5 days, s.c.) and ghrelin plus tilmicosin group (pretreatment with ghrelin followed by tilmicosin treatment). The heart were excised for evaluating Malondialdehyde (MDA) content, Superoxide Dismutase (SOD), Catalase (CAT) and Glutathione Peroxidase (GPx) activity. The results showed that tilmicosin treatment alone significantly (p<0.05) elevated the levels of MDA and lowered the activities of SOD, CAT and GPx when compared with the control group. Pretreatment with ghrelin ameliorated the SOD, CAT and GPx activities and inhibited the levels of MDA production in the heart tissue compared to tilmicosin-treated rats. The results of the study provide evidence that the ghrelin pretreatment enhances the antioxidant defense against tilmicosin-induced myocardial oxidative injury in rats and exhibit cardioprotective property.