In vitro assessment of the ethanol-induced hepatotoxicity on HepG2 cell line.
This study reports on measurement of the ethanol-induced hepatotoxicity in vitro on using the human hepatocyte cell line HepG2. Cells were incubated in the presence of increasing ethanol concentrations (10-80 mM). Cytotoxicity was quantitated spectrophotometrically both by the metabolism of the tetrazolium dye MTT and by the release into the medium of LDH and other marker enzymes of ethanol damage (AST, GGT and GHD). No cytotoxicity was observed up to 40 mM ethanol whereas a dose-dependent increment was found at higher concentrations (60-80 mM ethanol). At 80 mM ethanol, cell viability after 24 hours was reduced to 68% and 60% as assessed by MTT and LDH release respectively (p < 0.0001 vs. controls). Exposure for additional 24 hours did not increase cytotoxicity. Transmission electron microscopy revealed the presence of fat droplets (steatosis) and mitochondrial damage. The method reported appears to be an useful and reproducible technique for the in vitro assessment of the ethanol-induced cytotoxicity in a human liver cell line.
Neuman
M G
MG
Division of Clinical Nutrition, Hospital for Sick Children, Toronto, Ontario, Canada.
Koren
G
G
Tiribelli
C
C
D000704:Analysis of Variance; D001219:Aspartate Aminotransferases / Q000032:analysis; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000633:toxicity*; D005969:Glutamate Dehydrogenase / Q000032:analysis; D006801:Humans; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008099:Liver / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D008854:Microscopy, Electron; D008871:Microvilli / Q000187:drug effects / Q000648:ultrastructure; D008930:Mitochondria, Liver / Q000187:drug effects / Q000648:ultrastructure; D013778:Tetrazolium Salts; D013844:Thiazoles; D005723:gamma-Glutamyltransferase / Q000032:analysis
D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D000431:Ethanol; D007770:L-Lactate Dehydrogenase; D005969:Glutamate Dehydrogenase; D005723:gamma-Glutamyltransferase; D001219:Aspartate Aminotransferases; C022616:thiazolyl blue