Disulfiram may mediate erythrocyte hemolysis induced by diethyldithiocarbamate and 1,4-naphthoquinone-2-sulfonate.
The increase in 1,4-naphthoquinone-2-sulfonate (NQS)-induced hemolysis by the superoxide dismutase inhibitor diethyldithiocarbamate (DEDC) was formerly attributed to increased superoxide anion levels in the erythrocyte. Our results show that removal of DEDC after preincubation and prior to the addition of NQS did not produce a significant increase in hemolysis, which suggests that hemolysis is primarily caused by the reaction products of DEDC with NQS and not to the inactivation of superoxide dismutase. Disulfiram, the oxidized product of DEDC, was found to be the main product formed when excess DEDC was reacted with NQS. Oxygen uptake also occurred and hydrogen peroxide was formed. The latter caused the oxidation of DEDC to disulfiram as catalase prevented disulfiram formation. Disulfiram was found to readily hemolyze erythrocytes at low concentrations as well as to crosslink the proteins in the erythrocyte membrane. Furthermore, disulfiram-induced hemolysis was markedly enhanced in glutathione-depleted erythrocytes. Disulfiram was subsequently found to readily oxidize glutathione in red blood cells. When equimolar concentrations of DEDC and NQS were reacted, the major product formed was the diethyldithiocarbamate:1,4-naphthoquinone (DEDC:NQS) conjugate. However, the principal mediator of erythrocyte hemolysis when excess DEDC is reacted with 1,4-naphthoquinone-2-sulfonate is disulfiram, whose mode of action may be to modify membrane protein sulfhydryls.
Lauriault
V V
VV
Faculty of Pharmacy, University of Toronto, Ontario, Canada.
O'Brien
P J
PJ
D000818:Animals; D003432:Cross-Linking Reagents; D004221:Disulfiram / Q000494:pharmacology*; D004050:Ditiocarb / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000187:drug effects*; D005978:Glutathione / Q000097:blood; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D009285:Naphthoquinones / Q000494:pharmacology*; D010101:Oxygen Consumption; D051381:Rats; D011919:Rats, Inbred Strains; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors
D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D009285:Naphthoquinones; C019020:1,4-naphthoquinone-2-sulfonate; D004050:Ditiocarb; D013482:Superoxide Dismutase; D005978:Glutathione; D004221:Disulfiram