Ethanol induces the expression of alpha 1(I) procollagen mRNA in a co-culture system containing a liver stellate cell-line and freshly isolated hepatocytes.
To study the fibrogenic action of ethanol in vitro we used a co-culture system of freshly isolated hepatocytes and a liver stellate cell line (CFSC-2G) developed in our laboratory. Our results show that in this co-culture system ethanol induces the expression of alpha 1(I) procollagen mRNA in a dose- and time-dependent manner. This effect of ethanol was due to its metabolism by alcohol dehydrogenase since 4-methylpyrazole prevented the ethanol-mediated increase in alpha 1(I) procollagen mRNA. Ethanol was more efficient than acetaldehyde in inducing alpha 1(I) procollagen mRNA expression and its effect was protein synthesis-independent. Transfection of either hepatocytes or liver stellate cells with a reporter gene, chloramphenicol acetyl transferase (CAT), driven by 3700 bp of the mouse alpha 1(I) procollagen promoter demonstrated that only LSC expressed significant CAT activity and that this activity was enhanced by ethanol. Overall, our results suggest that this co-culture system is a useful model to study alcohol-induced fibrogenesis in vitro and that mechanisms other than acetaldehyde formation may also play an important role in alcohol-induced fibrogenesis.
Fontana
L
L
Division of Gastroenterology, Hepatology and Nutrition, Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Jerez
D
D
Rojas-Valencia
L
L
Solís-Herruzo
J A
JA
Greenwel
P
P
Rojkind
M
M
D000079:Acetaldehyde / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D000431:Ethanol / Q000494:pharmacology*; D005353:Fibronectins / Q000235:genetics; D000077604:Fomepizole; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D011347:Procollagen / Q000235:genetics*; D011720:Pyrazoles / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic / Q000187:drug effects
D005353:Fibronectins; D011347:Procollagen; D011720:Pyrazoles; D012333:RNA, Messenger; D000431:Ethanol; D000077604:Fomepizole; D000079:Acetaldehyde