Cardio-toxicity is considered one of the major concerns and drug-drug interactions could augment cardio-toxicities. Pioglitazone, an antidiabetic drug, had anticipations around its implications in cardiotoxicity. The antifungals fluconazole and itraconazole differ markedly in their potential to cause clinically significant drug interactions. The present study aimed to determine and compare the cardiotoxic potential of pioglitazone, fluconazole, itraconazole and their combined administration in adult healthy rats. Adult male Sprague Dawely rats were randomly and equally assigned into six groups of 10 rats per group. The first group was kept as control and received the vehicle. Three groups of animals were received single treatment, either pioglitazone (10 mg/kg), fluconazole (20 mg/kg) or itraconazole (18 mg/kg). Animal of the fifth group were received both pioglitazone (10 mg/ kg) and fluconazole (20 mg/kg) treatment, while the sixth group received combined pioglitazone (10 mg/kg) and itraconazole treatment (18 mg/kg). All treatments were administered orally for 28 consecutive days. At the end of experiment, electrocardiographs (ECG) were performed under anesthesia. Levels of cardiac biomarkers, serum Creatine kinase-MB (CK-MB), and troponin T (cTnT) levels were estimated. Oxidative stress parameters; catalase activity (CAT), lipid peroxidation (MDA) and reduced glutathione (GSH) were determined in cardiac tissue. Histological examination and immunohistochemical detection of cardiac caspase-3 reactivity were performed. Results revealed that pioglitazone and azole antifungals had cardiotoxic potential either as single treatment or in concomitant use. Treatments induced cardiotoxicity which directly affecting cardiomyocytes. Moreover, combined treatments induced more pronounced cardiac electrophysiological toxic effect. Hence, the therapeutic drug monitoring is essential.