Subnormothermic perfusion in the isolated rat liver preserves the antioxidant glutathione and enhances the function of the ubiquitin proteasome system

Abstract

The reduction of oxidative stress is suggested to be one of the main mechanisms to explain the benefits of subnormothermic perfusion against ischemic liver damage. In this study we investigated the early cellular mechanisms induced in isolated ratliversafter15minperfusionattemperaturesrangingfromnormothermia(37 ∘ C) to subnormothermia (26 ∘ Cand22 ∘ C). Subnormothermic perfusion was found to maintain hepatic viability. Perfusion at 22 ∘ C raised reduced glutathione levels and the activity of glutathione reductase; however, lipid and protein oxidation still occurred as determined by malondialdehyde, 4-hydroxynonenal-protein adducts, and advanced oxidation protein products. In livers perfused at 22 ∘ C the lysosomal and ubiquitin proteasome system (UPS) were both activated. The 26S chymotrypsin-like ( 훽 5) proteasome activity was significantly increased in the 26 ∘ C (46%) and 22 ∘ C (42%) groups. The increased proteasome activity may be due to increased Rpt6 Ser120 phosphorylation, which is known to enhance 26S proteasome activity. Together, our results indicate that the early events produced by subnormothermic perfusion in the liver can induce oxidative stress concomitantly with antioxidant glutathione preservation and enhanced function of the lysosomal and UPS systems. Thus, a brief hypothermia could trigger antioxidant mechanisms and may be functioning as a preconditioning stimulus.