Moderate alcohol consumption has been shown to reduce the morbidity and mortality from coronary heart disease. Ethanol elicits its protective effects via mechanisms that include activation of protein kinases linked to growth and survival. Our results in isolated neonatal rat cardiomyocytes demonstrate that repeated, short-term, low-dose exposure to ethanol is sufficient to activate the growth/survival pathways involving PKC, Akt and AMPK. Additionally, we are able to induce apoptosis in these cardiomyocytes using the saturated fatty acid, palmitate. Pretreatment with multiple low-dose ethanol exposures attenuates the apoptotic response to palmitate. This protection is manifested by a reduction in caspase-3-like activity, decreased mitochondrial loss of cytochrome c, and decreased loss of the mitochondrial lipid, cardiolipin. We have previously reported that incubation of cardiomyocytes with palmitate results in decreased production of ROS when compared to cells incubated with the non-apoptotic fatty acid oleate. In this present study, we observed an increase in the production of superoxide and the rates of fatty acid oxidation in cardiomyocytes pretreated with ethanol then exposed to fatty acids. The level of superoxide production in palmitate-treated cells returns to the levels observed in oleate-treated cells after ethanol exposure. Taken together with our observed increase in AMPK activity, we propose that ethanol pretreatments stimulate oxidative metabolism and electron transport within the cardiomyocyte. We postulate that the stimulation of palmitate metabolism may protect the cardiomyocyte by preventing the accumulation of unsaturated precursor molecules of cardiolipin synthesis. Maintaining cardiolipin levels may be sufficient to prevent the mitochondrial loss of cyt c and the downstream activation of caspases.