Molecular mechanisms of chloroacetaldehyde-induced cytotoxicity in isolated rat hepatocytes.
2-Chloroacetaldehyde (CAA) induced a loss in hepatocyte viability in a concentration- and time-dependent manner. Three phases before cytotoxicity ensued could be distinguished. Glutathione (GSH) was depleted immediately upon addition of CAA but only partial depletion occurred with subtoxic CAA concentrations. GSH-depleted hepatocytes were much more susceptible to CAA toxicity, indicating that CAA was detoxified by GSH. The second phase of changes involved a steady decrease in protein thiol levels, mitochondrial respiration, transmembrane potential and ATP levels. The third phase involved lipid peroxidation which commenced at around 60 min with a CAA concentration that caused 50% cytotoxicity in 120 min. Addition of antioxidants (diphenylphenylenediamine, butylated hydroxyanisole) and iron chelators (desferoxamine) at 40 min prevented lipid peroxidation and delayed CAA-induced cytotoxicity without restoring protein thiols, hepatocyte respiration or preventing further ATP depletion. Addition of dithiothreitol at 40 min, however, restored protein thiols and hepatocyte respiration, and prevented further ATP depletion and cytotoxicity. CAA-induced hepatocyte cytotoxicity therefore involved reversible thiol protein adduct formation, mitochondrial toxicity and lipid peroxidation.
Sood
C
C
Faculty of Pharmacy, University of Toronto, Ontario, Canada.
O'Brien
P J
PJ
D000079:Acetaldehyde / Q000031:analogs & derivatives* / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000172:deficiency; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D004229:Dithiothreitol; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000172:deficiency; D015227:Lipid Peroxidation; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008315:Malondialdehyde / Q000032:analysis; D010101:Oxygen Consumption / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D000975:Antioxidants; D008315:Malondialdehyde; D000255:Adenosine Triphosphate; C004656:chloroacetaldehyde; D005978:Glutathione; D000079:Acetaldehyde; D002118:Calcium; D004229:Dithiothreitol