Alcohol consumption for long periods negatively influences physiological functions of many cells, and leads to organ damage. Reactive oxygen and nitrogen species produced by ethanol metabolism cause adverse effects that might be alleviated by simultaneous treatment with various antioxidants. Here, the ability of ethyl pyruvate (EP) to reduce ethanol-induced oxidative stress was evaluated. Chemiluminescence studies show that EP has a higher capacity than pyruvate to scavenge hydrogen peroxide and superoxide anions. In order to evaluate whether EP can exert a protective effect against ethanol, rats were offered 10% ethanol in drinking burettes, containing or not different concentrations of EP (0.3%, 1% and 3%). The comet assay was employed to quantify the alcohol-induced DNA damage in rat lymphocytes. This test is a promising tool for the estimation of DNA damage at the single cell level. A significant protective effect of EP was observed in rat groups treated with this antioxidant, compared with those drinking only ethanol. Since EP has been shown to decrease the expression of numerous pro-inflammatory mediators, the monocyte respiratory burst was evaluated. The activation of monocyte NADPH oxidase by phorbol esters (PMA) showed that superoxide anion production was higher in the ethanol group than in the control group. The presence of EP considerably reduced superoxide anion production. In conclusion, hypotheses on possible mechanisms of action of EP on rat white blood cells are proposed.