Congenital heart defects affect 1% of born babies. Although corrective surgery is generally successful, there is an increased risk of morbidity and mortality in cyanotic children compared with acyanotic. PKC alpha is a member of the PKC family that is required for maintaining cell survival and cardiac function in the postnatal animal heart. Our previous data showed that PKCα mRNA expression is altered in cyanotic patients. However, little is known about human PKCα expression in response to chronic hypoxia, and its potential role in survival and resistance to reperfusion injury remains unexplored. We first investigated the effects of cyanosis on protein levels of PKCα and its cell survival targets eNOS and Bcl-2 in myocardium samples from paediatric patients suffering from Tetralogy of Fallot. Our findings showed an upregulation of PKCα protein expression by cyanosis that was associated with increased phosphorylated Bcl2 and eNOS levels in the myocardium. We then used the H9c2 myoblast cell line to test whether over-expressing PKCα results in increased levels of phosphorylated Bcl2 and eNOS. Infecting H9c2 cells with an adenovirus expressing a wild type form of PKCα resulted in an upregulation of phosphorylated-Bcl2 but not eNOS suggesting that PKCα survival effect is mediated through the anti-apoptotic protein Bcl2. A dominant negative form of PKCα did not alter levels of these two proteins. A potent PKC modulator, Bryostatin-1, that has been used in Alzheimer’s disease and cancer trials upregulated the phosphorylated PKCα protein levels within minutes to an hour in our cell model suggesting that this drug could be used in a cardiac surgery setting to activate PKCα. We finally tested the effect of this drug on a rat whole heart ischaemia/reperfusion injury model. Bryostatin-1 increased the functional recovery and reduced the infract size in rat heart subjected to ischaemia/reperfusion. In conclusion, our findings suggest that modulating PKCα provides a strategy for cardioprotection in patients with congenital heart defect undergoing corrective surgery.