Steatosis Reversibly Increases Hepatocyte Sensitivity to Hypoxia-Reoxygenation Injury

Abstract

Background: Steatosis decreases survival of liver grafts after transplantation due to poorly understood mechanisms. We examined the effect of steatosis on the survival of liver grafts in a rat liver transplantation model and the viability of cultured rat hepatocytes after hypoxia and reoxygenation. Materials and methods: Rats were fed a choline and methionine-deficient diet to induce hepatic steatosis, and the livers were transplanted into recipient rats after 6 h of cold storage. Cultured hepatocytes were made steatotic by incubation for 3 d in fatty acid-supplemented medium. Hypoxia and reoxygenation were induced by placing the cultures in a 90% N2/10% CO2 atmosphere for 4 h, followed by return to normoxic conditions for 6 h. Hepatocyte viability was assessed by lactate dehydrogenase release and mitochondrial potential staining. Results: Transplanted steatotic livers exhibited 0% viability compared with 90% for lean liver controls. When donor choline and methionine-deficient diet rats were returned to a normal diet, hepatic fat content decreased while viability of the grafts after transplantation increased. Cultured steatotic hepatocytes generated more mitochondrial superoxide, exhibited a lowered mitochondrial membrane potential, and released significantly more lactate dehydrogenase after hypoxia and reoxygenation than lean hepatocyte controls. When steatotic hepatocytes were defatted by incubating in fatty acid-free medium, they became less sensitive to hypoxia and reoxygenation as the remaining intracellular triglyceride content decreased. Conclusions: Hepatic steatosis reversibly decreases viability of hepatocytes after hypoxia and reoxygenation in vitro. The decreased viability of steatotic livers after transplantation may be due to a direct effect of hypoxia and reoxygenation on hepatocytes, and can be reversed by defatting. © 2009 Elsevier Inc. All rights reserved.