Tilmicosin is amacrolide antibiotic known to induce cardiotoxic effect when administered at large doses. The present study was carried out to investigate whether Lycopene (LYC) would ameliorate the acute cardiotoxic effect of tilmicosin antibiotic in treated rats. Fifty male albino rats were used throughout the experiment. They were divided equally into five groups, as follows: Group (1) (control), injected s/c with isotonic saline solution, Group (2) administrated 0.2 cm olive oil oral by stomach tube daily and kept also as control and scarified after 15 days. Group (3) administrated 10 mg / kg of lycopene dissolved in olive oil and given by stomach tube daily and scarified after 15 days Group (4) administrated 60 mg / kg of tilmicosin single subcutaneous dose and scarified after 5 days. Group (5) (Prophylactic group) administrated 10 mg / kg of lycopene for 15 days then 60 mg / kg tilmicosin s/c and scarified after 5 days. The effects of tilmicosin were evaluated with respect to alterations in serum, lactate dehydrogenase (LDH), creatine kinase (CK) activities also measurement of total malondialdehyde (MDA), catalase activity (CAT) and glutathione content of the heart tissues and histopathological findings of the heart sections in tilmicosin-treated rats with or without Lycopene. On the other hand, pretreatment of rats with Lycopene revealed marked decrease in cardiac biochemical parameters toward the normal limits. It concluded that lycopene effectively combated oxidative damage and protected antioxidant defense status of the cell against tilmicosin cardiotoxicity.