The potential use of cultured hepatocytes in predicting the hepatotoxicity of xenobiotics.
1. A protocol is proposed for screening for hepatotoxicity of xenobiotics in vitro in which hepatocytes exposed to the compounds are evaluated for both cytotoxic and metabolic effects. Four established hepatotoxins have been studied. 2. alpha-Amanitin at 1.5 pg/mg cell protein inhibited RNA synthesis by 93% and reduced albumin synthesis to 56% of the control after 13 h treatment. 3. D-Galactosamine at 40 microM inhibited glycogen synthesis by 31%, glucuronidation of p-nitrophenol by 13% and albumin synthesis by 10%, and produced an increase in cytosolic enzyme leakage. 4. Thioacetamide decreased ureogenesis after 24 h of treatment at 230 microM (31% inhibition) and after 48 h at 2.3 microM (25% inhibition). 5. Ultrastructural alterations of hepatocytes were found after 48 h exposure to 1 mM acetaminophen and were preceded by extensive leakage of the enzymes GOT and LDH. Membrane damage was observed after 24 h exposure to 0.1 mM acetaminophen.
Centro de Investigación, Hospital La Fe, Valencia, Spain.
M J
MJ
Gómez-Lechón
A
A
Montoya
P
P
López
T
T
Donato
A
A
Larrauri
J V
JV
Castell
D000081:Acetamides / Q000633:toxicity*; D000082:Acetaminophen / Q000633:toxicity*; D000546:Amanitins / Q000633:toxicity*; D000818:Animals; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005688:Galactosamine / Q000633:toxicity*; D007700:Kinetics; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D008854:Microscopy, Electron; D009963:Orotic Acid / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013853:Thioacetamide / Q000633:toxicity*
D000081:Acetamides; D000546:Amanitins; D013853:Thioacetamide; D000082:Acetaminophen; D009963:Orotic Acid; D005688:Galactosamine