Role of vitamin E in glutathione-induced oxidant stress: methemoglobin, lipid peroxidation, and hemolysis.
Red blood cells (RBC) from normal and vitamin E-deficient rats were incubated in a hypertonic solution of reduced glutathione adjusted to pH 8. Methemoglobin formation occurred in intact RBC from both normal and vitamin E-deficient rats. Hemolysis was significantly greater in RBC from vitamin E-deficient rats. Experiments with catalase, superoxide dismutase, and methional showed that H(2)O(2) was the primary extracellular source of oxidant stress. Extracellular superoxide and hydroxyl radical were not involved in oxidant stress. Experiments with dimethyl sulfoxide showed that intracellular hydroxyl radical, generated from H(2)O(2), was the hemolytic agent. Neither methemoglobin formation nor lipid peroxidation involved hydroxyl radical. Indeed, lipid peroxidation and hemolysis in RBC from vitamin E-deficient rats were concurrent rather than consecutive events. Phase contrast microscopy showed that rigid, crenated RBC with a precipitate around the interior periphery formed during glutathione-induced oxidant stress. The precipitate dissolved slowly as the crenated RBC were converted to smooth ghosts. It appeared that protein precipitates involving mixed disulfide bonds were reduced and solubilized when extracellular glutathione penetrated the ruptured cell. Comparisons between normal RBC and vitamin E-deficient RBC suggest that vitamin E has little effect on the inward diffusion of extra-cellular H(2)O(2). Vitamin E apparently interacts with different oxidant species derived from intracellular H(2)O(2) in preventing lipid peroxidation and the sulfhydryl group oxidation leading to hemolysis.
D000818:Animals; D002374:Catalase / Q000494:pharmacology; D002849:Chromatography, Gas; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005978:Glutathione / Q000494:pharmacology*; D006461:Hemolysis / Q000187:drug effects; D008055:Lipids / Q000097:blood*; D008297:Male; D008706:Methemoglobin / Q000378:metabolism*; D008858:Microscopy, Phase-Contrast; D010101:Oxygen Consumption / Q000187:drug effects; D010544:Peroxidases / Q000097:blood*; D051381:Rats; D013482:Superoxide Dismutase / Q000494:pharmacology; D014811:Vitamin E Deficiency / Q000097:blood*
D008055:Lipids; D008706:Methemoglobin; D010544:Peroxidases; D002374:Catalase; D013482:Superoxide Dismutase; D005978:Glutathione; D004121:Dimethyl Sulfoxide