Background: Isoflurane has a pharmacological precondition effect against ischemia injury in the heart, kidney and brain, but whether and how isoflurane precondition protects livers against ischemia and reperfusion(IR) injury is unclear.

Methods: Mice were randomly divided into isoflurane precondition(ISO) group and control group, receiving 1.5% isoflurane or carrier gas for 40 minutes respectively. A liver partial warm IR model was conducted and liver injury was evaluated. Primary hepatocytes were pretreated with 1.5% isoflurane for 2 hours prior to cell death induction by hydrogen peroxide. Cell death and survival were evaluated by LDH and CCK8 assay. Autophagy and regulatory molecules in stressed livers and hepatocytes were analyzed by western blot. Autophagy inhibitor(3-methyladenine,3-MA) and 5' adenosine monophosphate-activated protein kinase (AMPK) inhibitor(dorsomorphin) were administrated in vivo and in vitro.

Results: As compared to control group, mice in ISO group showed reduced liver injury and enhanced hepatocellular anti-apoptosis in livers post IR. Furthermore, liver autophagy was restored by ISO as indicated by elevated LC3B II protein level accompany with increased p62 degradation. In vitro study of primary hepatocytes also found that ISO effectively attenuated hepatocyte cell death induced by hydrogen peroxide. In addition, 3-MA pretreatment showed no significant influence in control group, but abrogated the protective role of ISO both in stressed livers and hepatocytes. Finally, signaling pathway analysis demonstrated that AMPK was activated by ISO. Pretreatment of AMPK inhibitor also abrogated liver protection by ISO, with no significant effect in control mice.

Conclusions: Our results indicate that isoflurane precondition attenuates liver IR injury via AMPK/mTOR-mediated hepatocellular autophagy restoration. Our findings provide a novel potential therapeutic strategy for managing liver IR injury.

CITATION INFORMATION: Rao Z, Zhou H, Pan X, Gao M, Wang Z, Sun J, Liu C, Ding Z. Isoflurane Precondition Alleviated Murine Liver Ischemia and Reperfusion Injury by Restoring AMPK/mTOR-Mediated Autophagy. Am J Transplant. 2017;17 (suppl 3).

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