title: Rat hepatocytes isolated from alcohol-induced fatty liver have an increased sensitivity to anoxic injury.
abstract: The aim of this study was to determine whether rat hepatocytes isolated from steatotic or nonsteatotic livers have different thresholds for injury due to anoxia-reoxygenation. Rats were fed ethanol or control diets for 8 weeks. Histology showed that more than 75% of the hepatocytes in alcohol-fed and less than 3% in control animals contained fatty vacuoles. The glycogen content was significantly reduced in steatotic livers. Isolated hepatocytes were cast in agarose gel threads and perfused with Krebs-Henseleit bicarbonate buffer. Cell viability was determined by Trypan Blue (TB) exclusion; cell injury was determined by lactate dehydrogenase (LDH) release; and superoxide anion (O2.-) was determined by lucigenin-enhanced chemiluminescence (LCL). During the pre-anoxic basal perfusion the following occurred: viability was 86% +/- 1% and 85% +/- 1%; LDH release was 16 +/- 3 and 15 +/- 3 mU/min; and LCL was 4 +/- 1 and 5 +/- 1 nA in steatotic and nonsteatotic hepatocytes, respectively. Cell viability decreased slightly under 4 hours of aerobic perfusion without differences between the two groups. In contrast, fatty hepatocytes died much faster than did control hepatocytes during anoxia; after 3 hours viability was 17% +/- 8% vs. 60% +/- 2% (P < .001), respectively. With reoxygenation following 2 hours of anoxia, the changes in viability, in LDH release, and in LCL were similar in both groups. These results indicate that in hepatocytes isolated from alcohol-fed rats when compared with control hepatocytes: 1) cell viability under aerobic conditions is not influenced; 2) anoxic injury is significantly increased; 3) a reduction in the hepatic glycogen stores, which may contribute to the enhanced sensitivity to anoxia, can be demonstrated; and 4) O2.- generation and cell injury occurring immediately after reoxygenation do not appear to be affected.
authors.affiliation: Department of Alcohol and Liver Disease, Oklahoma Medical Research Foundation, Oklahoma City, USA.
authors.forename: P
authors.initials: P
authors.lastname: Caraceni
authors.forename: H S
authors.initials: HS
authors.lastname: Ryu
authors.forename: V
authors.initials: V
authors.lastname: Subbotin
authors.forename: N
authors.initials: N
authors.lastname: De Maria
authors.forename: A
authors.initials: A
authors.lastname: Colantoni
authors.forename: L
authors.initials: L
authors.lastname: Roberts
authors.forename: F
authors.initials: F
authors.lastname: Trevisani
authors.forename: M
authors.initials: M
authors.lastname: Bernardi
authors.forename: D H
authors.initials: DH
authors.lastname: Van Thiel
mesh_terms: D000818:Animals; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology*; D000431:Ethanol / Q000008:administration & dosage / Q000633:toxicity; D005235:Fatty Liver, Alcoholic / Q000378:metabolism* / Q000473:pathology; D066298:In Vitro Techniques; D008099:Liver / Q000293:injuries* / Q000378:metabolism / Q000473:pathology; D008112:Liver Glycogen / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013481:Superoxides / Q000378:metabolism; D014617:Vacuoles / Q000187:drug effects / Q000473:pathology
chemical_list: D008112:Liver Glycogen; D013481:Superoxides; D000431:Ethanol