Rat hepatocytes isolated from alcohol-induced fatty liver have an increased sensitivity to anoxic injury.
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.
Caraceni
P
P
Department of Alcohol and Liver Disease, Oklahoma Medical Research Foundation, Oklahoma City, USA.
Ryu
H S
HS
Subbotin
V
V
De Maria
N
N
Colantoni
A
A
Roberts
L
L
Trevisani
F
F
Bernardi
M
M
Van Thiel
D H
DH
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
D008112:Liver Glycogen; D013481:Superoxides; D000431:Ethanol