An in vitro method of alcoholic liver injury using precision-cut liver slices from rats.
Alcohol abuse results in liver injury, but investigations into the mechanism(s) for this injury have been hampered by the lack of appropriate in vitro culture models in which to conduct in depth and specific studies. In order to overcome these shortcomings, we have developed the use of precision-cut liver slices (PCLS) as an in vitro culture model in which to investigate how ethanol causes alcohol-induced liver injury. In these studies, it was shown that the PCLS retained excellent viability as determined by lactate dehydrogenase and adenosine triphosphate (ATP) levels over a 96-h period of incubation. More importantly, the major enzymes of ethanol detoxification; alcohol dehydrogenase, aldehyde dehydrogenase, and cytochrome P4502E1, remained active and PCLS readily metabolized ethanol and produced acetaldehyde. Within 24 h and continuing up to 96h the PCLS developed fatty livers and demonstrated an increase in the redox state. These PCLS secreted albumin, and albumin secretion was decreased by ethanol treatment. All of these impairments were reversed following the addition of 4-methylpyrazole, which is an inhibitor of ethanol metabolism. Therefore, this model system appears to mimic the ethanol-induced changes in the liver that have been previously reported in human and animal studies, and may be a useful model for the study of alcoholic liver disease.
Klassen
Lynell W
LW
Experimental Immunology Laboratory, University of Nebraska Medical Center, Department of Internal Medicine, Section of Rheumatology, 983025 Nebraska Medical Center, Omaha, NE 68198-3025, USA.
Thiele
Geoffrey M
GM
Duryee
Michael J
MJ
Schaffert
Courtney S
CS
DeVeney
Amy L
AL
Hunter
Carlos D
CD
Olinga
Peter
P
Tuma
Dean J
DJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D000426:Alcohol Dehydrogenase / Q000378:metabolism; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism; D004195:Disease Models, Animal; D000431:Ethanol / Q000633:toxicity*; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D008108:Liver Diseases, Alcoholic / Q000201:enzymology / Q000209:etiology / Q000473:pathology; D008297:Male; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar; D014280:Triglycerides / Q000378:metabolism
D014280:Triglycerides; D000431:Ethanol; D000255:Adenosine Triphosphate; D000426:Alcohol Dehydrogenase; D019392:Cytochrome P-450 CYP2E1