Altered activities of antioxidant defence system enzymes and the levels of free radicals scavengers have been found to correlate with various physiological or pathological conditions, including stress. The aim of this study was to determine the effects of chronic 21 day isolation stress on antioxidant enzymes (AOEs) expression and activity in Wistar rat liver tissue. The serum corticosterone (CORT) and glucose (GLU) levels were also measured, as one of the most important indicators of stress. Our data revealed that in chronic stress conditions, when both CORT and GLU were low, the AOEs expression was markedly induced. This increase in MnSOD, CuZnSOD, and catalase exhibited similar trend implying efficient detoxification of O-2 radical anion and H2O2. However, this trend was not followed by the respective enzyme activity. While the total SOD activity was induced by the stress, catalase activity remained unaltered. This discrepancy led us to a conclusion that chronic isolation stress may cause oxidant-antioxidant imbalance in rat liver tissue, favoring H2O2 accumulation.