Molecular mechanisms of dibromoalkane cytotoxicity in isolated rat hepatocytes.
The cytotoxicity of dibromoalkanes to isolated hepatocytes was proportional to the dibromoalkane concentration and increasing chain length of the dibromoalkane (C2-C6). The rapid hepatocyte glutathione (GSH) depletion which occurred upon addition of the dibromoalkanes was also dependent on the concentration and chain length of the dibromoalkane. When added to hepatocytes, dibromoalkanes also caused a loss in protein sulfhydryl groups. After a lag period, lipid peroxidation occurred before the onset of cytotoxicity. Antioxidants or removing the oxygen from the medium markedly delayed dibromoalkane cytotoxicity. Bromoaldehydic metabolites formed by cytochrome P450-dependent mixed-function oxidases were probably responsible for lipid peroxidation as deuterated 1,2-dibromoethane (d4-DBE) induced less lipid peroxidation and was less cytotoxic even though GSH was depleted as rapidly and as effectively. Hepatocytes were also more resistant to dibromoalkanes if cytochrome P450 isoenzymes were inactivated with SKF 525A or methyl pyrazole. Furthermore, hepatocyte susceptibility to dibromoalkanes was increased markedly if aldehyde dehydrogenase was inactivated with disulfiram, cyanamide or chloral hydrate. Cytochrome P450-induced hepatocytes isolated from pyrazole-, phenobarbital- or 3-methylcholanthrene-pretreated rats were also more susceptible to dibromoalkanes. These results suggest that dibromoalkane-induced cell lysis is due to lipid peroxidation as well as cytochrome P450-dependent formation of toxic bromoaldehydic metabolites which can bind with cellular macromolecules. Dibromoethane GSH conjugates also contribute to DBE cytotoxicity as depleting hepatocyte GSH beforehand increased hepatocyte resistance to DBE but not other dibromoalkanes.
Khan
S
S
Faculty of Pharmacy, University of Toronto, Ontario, Canada.
Sood
C
C
O'Brien
P J
PJ
D000079:Acetaldehyde / Q000031:analogs & derivatives / Q000378:metabolism / Q000633:toxicity; D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors; D000818:Animals; D003484:Cyanamide / Q000494:pharmacology; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004221:Disulfiram / Q000494:pharmacology; D000431:Ethanol / Q000031:analogs & derivatives / Q000378:metabolism / Q000633:toxicity; D015946:Ethylene Dibromide / Q000378:metabolism / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism; D006842:Hydrocarbons, Brominated / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D015227:Lipid Peroxidation; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D008954:Models, Biological; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley
D065607:Cytochrome P-450 Enzyme Inhibitors; D006842:Hydrocarbons, Brominated; C038235:bromoacetaldehyde; D015946:Ethylene Dibromide; D000431:Ethanol; D003484:Cyanamide; D003577:Cytochrome P-450 Enzyme System; D000444:Aldehyde Dehydrogenase; D005978:Glutathione; D000079:Acetaldehyde; D004221:Disulfiram; C025475:ethylene bromohydrin