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Positive effects of calorie restrictions (CR) and EOD include decreased body weight, prolonged life span, but also changes in metabolism of the liver. In present paper our aim was to examine antioxidative enzymes: Catalase (CAT) and Manganese superoxidative dismutase (MnSOD, SOD2) and Glutathione peroxidase 4 (Gpx-4) in connection to caspase-3 and inflammatory mediators (IL-1β and TNF- α) in EOD liver tissue in comparison to control mice. After 9 months of EOD treatment male mouse liver tissue was harvested and prepared for analysis. Protein semi-quantitative estimation and cellular immunolocalization was performed for CAT, SOD2, Gpx-4, caspase-3, IL-1β and TNF- α in liver tissue of mice fed every-other day in comparison to control (AL fed) animals. After prolonged EOD feeding in mice we observed decreased level of SOD2 and Gpx-4, decreased caspase-3, IL-1β and TNF-α expression in liver tissue on protein level measured by semi-quantitative DAB staining intensity. For the first time we showed immunolocalization of major antioxidative enzymes (CAT, SOD2, Gpx-4) in liver tissue after DR. Decrease of two major antioxidant enzymes combined with decrease of apoptotic marker and inflammatory factors indicate decrease in oxidative stress as the result of fast in EOD feeding regime. • Dietary regimes influence oxidative stress level in liver. • Prolonged every-other day (EOD) feeding reduces SOD2 and GPX-4 in liver cells. • Prolonged EOD reduces caspase-3 and IL-1β in liver. • SOD2 and GPX-4 are expressed in hepatocytes, Kupffer cells and LSEC in normal liver. [ABSTRACT FROM AUTHOR]