Despite years of experimental and clinical research, myocardial ischemia-reperfusion (IR) remains an important cause of cardiac morbidity and mortality. The transcription factor nuclear factor-kappa B (NF-B) has been implicated as a key mediator of reperfusion injury. Activation of NF-B is dependent upon the phosphorylation of its inhibitor, IB, by the specific inhibitory kappa B kinase (IKK) subunit, IKK. We hypothesized that specific antagonism of the NF-B inflammatory pathway through IKK inhibition reduces acute myocardial damage following ischemia-reperfusion injury. C57BL/6 mice underwent left anterior descending (LAD) artery ligation and release in an experimental model of acute IR. Bay 65-1942, an ATP-competitive inhibitor that selectively targets IKK kinase activity, was administered intraperitoneally either prior to ischemia, at reperfusion, or 2 hours after reperfusion. Compared with untreated animals, mice treated with IKK inhibition had significant reduction in left ventricular infarct size. Cardiac function was also preserved following pre-treatment with IKK inhibition. These findings were further associated with decreased expression of phosphorylated-IB and phosphorylated-p65 in myocardial tissue. In addition, IKK inhibition decreased serum levels of TNF- and IL-6, two prototypic downstream effectors of NF-B activity. These results demonstrate that specific IKK inhibition can provide both acute and delayed cardioprotection and offers a clinically accessible target for preventing cardiac injury following ischemia-reperfusion.