Background: Cardiac-related problems accounts for about 10%–35% of all deaths. Medicinal plants and their derivatives are a rich source of effective cardioprotective agents. Hecogenin (HCG) is a sapogenin with a wide spectrum pharmacological property. In this study, we aimed to evaluate the cardioprotective effect of HCG against isoproterenol (ISO)-induced myocardial infarction (MI) in rats. Materials and Methods: The animals were divided into four groups. Group I: Control group consisted ofrats fed onstandard pellet (23 days); Group II: HCG group, HCG 50 μg/kg body weight (BW), oral administration, (P. O) on 21 days; Group III: ISO group, 60 mg ISO/kg BW; subcutaneous (sc) administered on days 22 and 23; Group IV: HCG (50 μg/kg BW. P. O) (21 days) + ISO (days 22 and 23). After sacrificing the animals, we analyzed thebiochemical and molecular markers. Results: Our results showed that there were no significant differences in the BW of rats; however, the HCG and ISO groups showed a significant reduction in heart weight. When compared with ISO group, hepatic and cardiac biomarkers were low in HCG + ISO group. Furthermore, compared to the ISO group, the level of lipid peroxidation products was restored to their optimal level in the HCG + ISO group. According to the histopathological findings, animals from HCG group demonstrated restoration of their tissue architecture. Immunohistochemistry demonstrated that animals in the HCG group showed reduced expression levels of nuclear factor kappa B (NF-κB) and p53, which was similar to the control and HCG group. This demonstrates cardioprotective effects of HCG. Conclusion: HCG attenuated ISO-induced MI via inhibition of activation of NF-κB and p53 signaling pathway. Overall, the findings recommended that HCG is a promising therapeutic agent for the treatment of MI.