Interaction of 1-hydroxyethyl radical with antioxidant enzymes.
There is considerable interest in the role of the 1-hydroxyethyl radical (HER) in the toxic effects of ethanol. The goal of this study was to evaluate the effects of HER on classical antioxidant enzymes. The interaction of acetaldehyde with hydroxylamine-o-sulfonic acid has been shown to produce 1, 1'-dihydroxyazoethane (DHAE); this compound appears to be highly unstable, and its decomposition leads to the generation of HER. Addition of DHAE into a solution of PBN led to the appearance of the typical EPR spectra of PBN/HER adduct. No PBN/HER spin adduct was detected when DHAE was incubated with 0.1 M PBN in the presence of GSH. In the absence of PBN, DHAE oxidized ascorbic acid to semidehydroascorbyl radical, presumably via an ascorbate-dependent one-electron reduction of HER back to ethanol. Catalase was progressively inactivated by exposure to DHAE-generated HER in a time and HER concentration-dependent manner. Ascorbic acid and PBN gave full protection to catalase against HER-dependent inactivation. The antioxidants 2-tert-butyl-4-methylphenol, propylgallate, and alpha-tocopherol-protected catalase against inactivation by 84, 88, and 39%, respectively. Other antioxidant enzymes were also sensitive to exposure to HER. Glutathione reductase, glutathione peroxidase, and superoxide dismutase were inactivated by 46, 36, and 39%, respectively, by HER. The results reported here plus previous results showing HER interacts with GSH, ascorbate, and alpha-tocopherol suggest that prolonged generation of HER in cells from animals chronically exposed to ethanol may lower the antioxidant defense status, thereby contributing to mechanisms by which ethanol produces a state of oxidative stress and produces toxicity.
Puntarulo
S
S
Physical Chemistry, School of Pharmacy and Biochemistry, University of Buenos Aires, Argentina.
Stoyanovsky
D A
DA
Cederbaum
A I
AI
D000975:Antioxidants / Q000378:metabolism*; D001205:Ascorbic Acid / Q000378:metabolism; D001391:Azo Compounds / Q000378:metabolism; D002084:Butylated Hydroxytoluene / Q000031:analogs & derivatives / Q000378:metabolism; D002374:Catalase / Q000037:antagonists & inhibitors / Q000378:metabolism; D004228:Dithionitrobenzoic Acid / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D000431:Ethanol / Q000378:metabolism* / Q000633:toxicity; D016166:Free Radical Scavengers / Q000378:metabolism; D005609:Free Radicals / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D005980:Glutathione Reductase / Q000037:antagonists & inhibitors / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects; D011435:Propyl Gallate / Q000378:metabolism; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors / Q000378:metabolism; D014810:Vitamin E / Q000378:metabolism
C403003:1,1'-dihydroxyazoethane; D000975:Antioxidants; D001391:Azo Compounds; D016166:Free Radical Scavengers; D005609:Free Radicals; D014810:Vitamin E; C065424:1-hydroxyethyl radical; D002084:Butylated Hydroxytoluene; D000431:Ethanol; D011435:Propyl Gallate; D004228:Dithionitrobenzoic Acid; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D005980:Glutathione Reductase; D005978:Glutathione; D001205:Ascorbic Acid; C054527:2-tert-butyl-4-methylphenol