Bromobenzene and furosemide hepatotoxicity: alterations in glutathione, protein thiols, and calcium.
The nature of the events whereby the reactive intermediates resulting from the bioactivation of bromobenzene and furosemide induce hepatotoxicity is unknown. To examine a role for disturbances in intracellular calcium homeostasis, secondary to a depletion in cellular reduced glutathione (GSH) and reduced protein thiols (PSHs), isolated mouse hepatocytes were exposed to cytotoxic concentrations of bromobenzene or furosemide. Cytosolic calcium concentration, as well as thiol status, was determined. The incubation of hepatocytes with 3.0 mM bromobenzene, and subsequent additions (1.2 mM) of the agent every hour, resulted in significant GSH depletion. The loss of plasma membrane integrity at 1.5 h preceded both a rise in the cytosolic Ca2+ concentration and depletion of total PSH content. Furosemide (1.0 mM) produced a 70% depletion in cellular GSH content in isolated hepatocytes. The initiation of cell damage occurred concurrently with both a rise in the cytosolic Ca2+ concentration and a depletion of total PSH content 4 h following furosemide addition. Since the increase in cytosolic Ca2+ did not precede cytotoxicity, these results do not support an initiating role for Ca2+ deregulation in bromobenzene and furosemide hepatotoxicities. In addition, depletion of PSH content did not correlate with bromobenzene- or furosemide-induced cytotoxicity.
Grewal
K K
KK
Department of Pharmacology and Toxicology, Queen's University, Kingston, ON, Canada.
Rafeiro
E
E
Racz
W J
WJ
D000818:Animals; D001969:Bromobenzenes / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology*; D003600:Cytosol / Q000378:metabolism; D004232:Diuretics / Q000633:toxicity*; D005665:Furosemide / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism*; D006706:Homeostasis / Q000187:drug effects; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D011506:Proteins / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D001969:Bromobenzenes; D004232:Diuretics; D011506:Proteins; D013438:Sulfhydryl Compounds; D005665:Furosemide; C032036:bromobenzene; D005978:Glutathione; D002118:Calcium