Ischemia-reperfusion (IR) injury is a major clinical problem and is associated with numerous adverse effects. GGsTop [2-amino-4{[3-(carboxymethyl)phenyl](methyl)phosphono}butanoic acid] is a highly specific and irreversible -glutamyl transpeptidase (-GT) inhibitor. We studied the protective effects of GGsTop on IR induced hepatic injury in rats. Ischemia was induced by clamping the portal vein and hepatic artery of left lateral and median lobes of the liver. Before clamping, saline (IR group) or saline containing 1 mg/kg body weight of GGsTop (IR-GGsTop group) was injected into the liver through inferior vena cava. At 90 min of ischemia, blood flow was restored. Blood was collected before induction of ischemia and prior to restoration of blood flow, and at 12, 24, and 48 h after reperfusion. All the animals were sacrificed at 48 h after reperfusion and the livers were harvested. Serum levels of ALT, AST, and -GT were significantly lower after reperfusion in IR-GGsTop group compared to IR group. Massive hepatic necrosis was present in IR group, while only few necroses were present in IR-GGsTop group. Treatment with GGsTop increased hepatic GSH content which was significantly reduced in IR group. Furthermore, GGsTop prevented increase of hepatic -GT, malondialdehyde, 4-hydroxynonenal, and TNF-α while all these molecules significantly increased in the IR group. In conclusion, treatment with GGsTop increased glutathione levels and prevented formation of free radicals in the hepatic tissue that lead to decreased IR-induced liver injury. GGsTop could be used as a pharmacological agent to prevent IR-induced liver injury and the related adverse events.