Development of an in vitro hepatotoxicity assay for assessing the effects of chronic drug exposure.
We have used the human hepatoma cell line HepG2 to compare the hepatotoxic effects of acute (24 hr) and chronic (up to 10 days) exposure to amitriptyline, paracetamol and ondansetron. In acute exposure studies, hepatotoxicity was assessed by the sulforhodamine B protein staining method, where amitriptyline and paracetamol produced 50% hepatotoxicity at concentration of 30 microM and 7 mM, respectively, while ondansetron was non-hepatotoxic at 100 microM, the highest concentration used. In chronic exposure studies, the morphology of HepG2 cells was assessed by phase microscopy every 2 days and the compounds, at concentrations determined from the acute assay, were added fresh every 2 days. Chronic exposure to amitriptyline and paracetamol produced significant morphological changes in HepG2 cells at 3 microM and 1 mM respectively, concentrations which had no significant effect in the acute assay. Ondansetron (100 microM) produced only slight morphological changes in the cells after 10 days of culture. The combination of acute and chronic drug exposure assays with HepG2 cells represents novel in vitro systems for the hepatotoxicological assessment of drugs intended for human use.
Hall
T J
TJ
MD Laboratories, Bartenheim, France.
James
P R
PR
Cambridge
G
G
D000082:Acetaminophen / Q000633:toxicity*; D000639:Amitriptyline / Q000633:toxicity*; D002460:Cell Line; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000473:pathology*; D008107:Liver Diseases / Q000175:diagnosis*; D017294:Ondansetron / Q000633:toxicity*
D000639:Amitriptyline; D000082:Acetaminophen; D017294:Ondansetron