The aim of the present study was to evaluate the efficacy of pre-supplementation with pomegranate (Punica granatum L.) juice (PJ) on heart weight, infarct size, plasma marker enzymes of cardiac damage, lipid peroxidation, endogenous enzymatic and non-enzymatic antioxidants, cardiac ATPases and histopathology of isoproterenol (IP)-induced cardiac necrosis (CN) in rats. Rats treated with IP (85 mg/kg, s.c.) for 2 days at an interval of 24 h caused significant (P < 0.05) infarct in myocardium and increase in heart weight, lipid peroxidation (LPO), activity levels of Ca(+2) ATPase and plasma marker enzymes, while there was significant (P < 0.05) decrease in endogenous enzymatic and non-enzymatic antioxidants and Na(+)-K(+) and Mg(+2)ATPases. Pre-supplementation with PJ for 30 consecutive days and treated with IP on days 29th and 30th showed significantly (P < 0.05) lesser increase in heart weight, infarct size, plasma marker enzymes, lipid peroxidation, Ca(+2) ATPase and a significant protective effect in endogenous enzymatic and non-enzymatic antioxidants, Na(+)-K(+) and Mg(+2) ATPases compared to IP alone treated group. Present study provides first scientific report on protective effect of supplementation of Pomegranate juice against IP-induced CN in rats.