During myocardial ischemia/reperfusion (I/R), the generation of reactive oxygen species (ROS) contributes to post‐reperfusion cardiac injury and contractile dysfunction. Activation of protein kinase C epsilon (PKC ɛ) during I/R has been shown to increase ROS release, in part, by its stimulation of increased uncoupled endothelial nitric oxide synthase activity. We hypothesize that using a cell permeable PKC ɛ peptide inhibitor (PKC ɛ‐) (N‐myr‐EAVSLKPT, MW=1054 g/mol, 10μM or 20μM) will improve post‐reperfused cardiac function and attenuate infarct size compared to untreated controls in isolated perfused rat hearts subjected to I(30 min)/R(90 min). Male Sprague‐Dawley rats (275–325 g) were anesthetized with sodium pentobarbital (60 mg/kg) and anticoagulated with heparin 1000 units IP. PKC ɛ‐was dissolved in Krebs' buffer and infused during the first 10 min of reperfusion. PKC ɛ‐treated hearts exhibited significant improvement in post‐reperfused cardiac function at 90 min in the maximal rate of left ventricular developed pressure (+dP/dtmax): 56±5%; n=6 (10μM) and 46±3%; n=4 (20μM) compared to untreated controls (n=6) which only recovered to 32±5% of baseline values for +dP/dtmax respectively (p<0.05). Furthermore, PKC ɛ‐treated hearts showed significant reduction in infarct size of 27±2% (10μM) and 28 ±2% (20μM) compared to untreated control I/R hearts, 40±3% (p<0.05). The results suggest that PKC ɛ‐is effective in improving cardiac function and reducing infarct size and is a putative treatment that could aid in clinical myocardial infarction/organ transplantation patient recovery.Support or Funding InformationThis study was supported by the Center for Chronic Disorders of Aging, the Division of Research and the Department of Bio‐Medical Sciences at Philadelphia College of Osteopathic Medicine.