The antidepressant-like effect of repeated administration of diphenyl diselenide (PhSe)2 in rats exposed to malathion is reported. The role of Na+K+ ATPase, acetylcholinesterase (AChE) and monoamine oxidase (MAO) activities and oxidative stress in antidepressant behavior were investigated in cerebral cortex of rats. Rats were exposed once a day for 3 consecutive days to malathion (50mg/kg, intraperitoneal) and (PhSe)2 (50mg/kg, oral). To investigate the antidepressant-like behavior rats were submitted to the forced swimming test (FST) and open-field test (OFT). Thiobarbituric acid reactive species (TBARS) levels, enzymatic and non-enzymatic antioxidant defenses were carried out in cerebral cortex of rats. The results confirmed that malathion increased immobility time in the FST without altering the locomotor performance in the OFT. Treatment with (PhSe)2 ameliorated performance in the FST without altering the crossing numbers in the OFT. The inhibition of Na+K+ ATPase activity caused by malathion was prevented by treatment with (PhSe)2. Exposure to malathion did not alter parameters of oxidative stress as well as AChE and MAO activities in cerebral cortex of rats. In conclusion, (PhSe)2 exerted antidepressant-like effect in rats exposed to malathion. Na+K+ ATPase activity is, at least in part, involved in (PhSe)2 antidepressant-like behavior.