Cytotoxicity of bile in human Hep G2 cells and in primary cultures of rat hepatocytes.
There has been increasing interest in the development of a hepatocyte bioreactor for the treatment of acute hepatic failure; however, little is known about the effect of hepatocyte byproducts on the viability of the cells in the bioreactor environment. We investigated the effects of increasing concentrations of bile on the growth and viability of the human hepatoma cell line Hep G2 and on the cytochrome P-450 content and dependent mixed function oxidase (MFO) activities, reduced glutathione (GSH) content, and glutathione S-transferase (GST) activity of primary cultures of rat hepatocytes. Our purpose was to determine whether or not it would be necessary to pretreat the plasma from patients with acute liver failure to remove elevated bile concentrations which might be toxic to the hepatocytes in an artificial liver device. Bile was found to inhibit Hep G2 cell growth at concentrations as low as 0.1% and to decrease viability at concentrations above 0.5%. The cytochrome P-450 and GSH contents and the activities of the MFO system and of GST were decreased in the primary cultures of hepatocytes following 24 h treatment with concentrations of bile at and above 0.5%. The MFO activities associated with different cytochrome P-450 isoenzymes decreased to different extents in the presence of bile with the O-dealkylation of pentoxyresorufin being more labile than that of ethoxyresorufin. Our data indicate that elevated bile concentrations are cytotoxic to liver cells, and it may be necessary to pretreat patient plasma to decrease its bile content to protect the cells during the clinical operation of a hepatocyte bioreactor device.
Smirthwaite
A D
AD
Bioengineering Unit, University of Strathclyde, Glasgow, Scotland.
Gaylor
J D
JD
Cousins
R B
RB
Grant
M H
MH
D000818:Animals; D001646:Bile / Q000737:chemistry* / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D003640:Dealkylation; D005978:Glutathione / Q000032:analysis; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000032:analysis; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D017114:Liver Failure, Acute / Q000097:blood / Q000503:physiopathology / Q000628:therapy; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D019164:Liver, Artificial; D008297:Male; D010078:Oxazines / Q000378:metabolism; D010088:Oxidoreductases / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D010078:Oxazines; C007649:ethoxyresorufin; C044943:pentoxyresorufin; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D005982:Glutathione Transferase; D005978:Glutathione