OBJECTIVES: This study aimed to investigate the potential protective effects of crocin against azithromycin (AZ)-induced cardiotoxicity. MATERIALS AND METHODS: The experimental design consisted of four groups: Control, crocin, Azithromycin, and crocin plus Azithromycin (AZ+CR 50). To evaluate oxidative stress, inflammation, and apoptosis in cardiac tissue, a combination of biochemical, molecular, and histological techniques was employed. Biomarkers such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), nuclear factor erythroid 2-related factor 2 (Nrf-2), and heme oxygenase-1 (HO-1) were assessed to determine anti-oxidant status, while malondialdehyde (MDA) and advanced oxidation protein products (AOPP) were measured as indicators of oxidative damage. Protein levels of inflammatory markers NF-κB and toll-like receptor 4 (TLR-4) and apoptotic regulators Bax, Bcl-2, and Caspase-3 were quantified. RESULTS: Crocin treatment effectively attenuated AZ-induced oxidative stress by enhancing anti-oxidant enzyme activity and reducing MDA and AOPP levels. Furthermore, crocin significantly down-regulated the expression of NF-κB and TLR-4 proteins, indicating reduced inflammation. The proapoptotic proteins Bax and Caspase-3, which were elevated following AZ exposure, were markedly decreased by crocin co-treatment. Conversely, the reduced expression of the antiapoptotic protein Bcl-2 caused by AZ was restored by crocin. In addition, AZ administration led to increased levels of COX-2 and MAPK-3, both of which were down-regulated following crocin treatment. Histological analysis revealed that crocin reduced degenerative and necrotic changes in heart tissue caused by AZ. CONCLUSION: These findings suggest that crocin exerts cardioprotective effects against AZ-induced damage by modulating oxidative stress, inflammation, and apoptosis.