Abstract Tilmicosin (Til) was purposed to be used in the treatment of a wide range of respiratory diseases in livestock. However, undesirable adverse effects, cardiac toxicity, in particular, may be associated with Til therapy. In the present study, the response of adult rats administered Til subcutaneously at different doses (10, 25, 50, 75, and 100 mg/kg b.w.; single injection) was evaluated. Astragalus polysaccharide (AP) at two doses (100 and 200 mg/kg b.w.; intraperitoneally) was investigated for its potential to counteract the cardiac influences, involving the oxidative stress-induced damage and apoptotic cell death, elicited by the Til treatment at a dose of 75 mg/kg b.w. in rats. Til induced mortalities and altered the levels of the biomarkers for the cardiac damage, particularly in the rats treated with the doses of 75 and 100 mg/kg b.w.; similarly, morphological alterations in cardiac tissue were seen at all studied doses. AP was found to cause a significant (P < 0.05) decline in the levels of impaired cardiac injury markers (troponin, creatine phosphokinase, and creatine phosphokinase-MB), improvement in the antioxidant endpoints (total antioxidant capacity), and attenuation in the oxidative stress indices (total reactive oxygen species, 8-hydroxy-2-deoxyguanosine, lipid peroxides [malondialdehyde], and protein carbonyl), associated with a significant (P < 0.05) modulation in the mRNA expression levels of the encoding genes (Bcl-2, Bax, caspase-3, P53, Apaf-1, and AIF), related to the intrinsic pathway of apoptotic cell death in the cardiac tissue. AP administration partially restored the morphological changes in the rat's heart. The highest protective efficacy of AP was recorded at a dose level of 200 mg/kg b.w. Taken together, these results indicated that AP is a promising cardioprotective compound capable of attenuating Til-induced cardiac impact by protecting the rat cardiac tissue from Til-induced apoptosis when administered concurrently with and after the Til injection.