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Experimental studies indicate that oxidative stress during and after laparoscopic surgery may cause liver ischemia-reperfusion injury. The aim of the study was to assess the effect of ischemic preconditioning against liver damage during pneumoperitoneum on oxidative stress. Twenty one New Zealand rabbits were divided into three groups of seven animals. Control group (C) rabbits received anesthesia for 60 min alone; 15 mm Hg intra-abdominal pressure with CO2 for 60 min was used in the pneumoperitoneum group animals (PNP); and 15-min insufflation and 10-min desuflation followed by 60-min pneumoperitoneum were used in the ischemic preconditioning group animals (IP). Venous blood samples were obtained at different time points to measure lipid hydroperoxide, glutathione reductase and total antioxidant status as indicators of increased oxidative stress. Aspartate transaminase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels were evaluated as indicators of hepatocellular injury. The Kruskal-Wallis and Mann-Whitney U tests were used on statistical analysis. Elevated intra-abdominal pressure was found to produce significant increase in lipid hydroperoxide at the end of pneumoperitoneum and 30 min after desuflation in comparison with pre-insufflation period, and with both C and IP groups at the same time points. Total antioxidant status level decreased significantly in the PNP group at 24 h of desuflation. At 24h of desuflation, the AST, ALT and LDH levels were significantly increased in the PNP group in comparison with the levels measured before induction of anesthesia, and with the C and IP groups. Study results demonstrated that ischemic preconditioning prevented hepatocyte injury and oxidative stress during CO2 pneumoperitoneum.