Background: Neuroprotection during cardiac and aortic arch surgeries has been a topic of extensive concern. Several protocols were developed in the trial to achieve the best neurological outcome; amongst them is nitric oxide (NO) supplementation through extracorporeal membrane oxygenation (ECMO). While NO was shown to influence the neurological outcome when supplemented during cardiac surgeries, a little is known about the cardiac effects. The current study addresses the molecular effects associated with NO supplementation on the myocardium. Methods: Male Sprague Dawley rats (n=20, 450-550gm) were anesthetized and cooled with cardiopulmonary bypass (CPB) to a core temperature of 19±1 o C and maintained in hypothermic cardiac arrest for 30 minutes. Then, rats were rewarmed to a target temperature of 35 o C for 60 min using CPB without (control group, CPB), or with NO 20 ppm supplementation (experimental group). By the end of experiment, hearts were harvested and analyzed. Results: RNA sequencing analysis showed NO supplementation results in the differential expression of more the genes related to lipid metabolism, oxidative phosphorylation, inflammatory signaling, proteasome formation, autophagy, calcium handling, smooth muscle relaxation, and apoptotic response compared to the control group. RT-PCR confirmed the RNA sequencing analysis. The experimental group showed a six-fold increase in inflammatory markers IL-1 and NF-κβ. The control group showed increased expression of α1 and β1 adrenergic receptors, NO treatment has led to normalization of expression and a three-fold increase in β2 receptors expression compared to shams. NO supplementation induced apoptosis in the myocardium, as indicated by a 50% increase in TUNEL-positive nuclei compared to the control group. Conclusions: NO supplementation during cardiac arrest boosts the inflammatory response in the myocardium and is associated with increased apoptosis.