Background: Hepatic ischemia reperfusion injury (HIRI) is considered one of the most common causes of liver damage and dysfunction. Oxytocin (OT), besides its classical functions, exhibits a potent antistress, anti-inflammatory, and antioxidant effects. Aim: This study was designed to evaluate the effect of pretreatment with OT on HIRI and to determine its possible protective mechanisms, focusing on the potential role of NADPH oxidase 2 (NOX2). Methods: 28 adult Wister albino male rats were divided into 4 groups: group I (control group): received saline and subjected to surgery without ischemic procedure, group II (OT group): received OT and subjected to surgery without ischemic procedure, group III (HIR group): underwent hepatic ischemia reperfusion (HIR) procedure & group IV (HIR+ OT group): received OT and underwent HIR procedure. We assessed the effect of OT on serum liver enzymes, hepatic malondialdehyde (MDA), glutathione (GSH), tumor necrosis factor alfa (TNFα), and NOX2. Results: HIR caused significant increase in serum liver enzymes, hepatic MDA, TNFα and NOX2 levels with a significant decrease in GSH level. Administration of OT caused a significant improvement in all previous parameters, these results were supported by histopathological examination. Conclusion: OT exerts hepatoprotective effect in HIR-induced liver injury even in part through NOX2.