Similarities in the metabolism of alloxan and dehydroascorbate in human erythrocytes.
The beta-cell toxin alloxan is reduced within cells to dialuric acid, which may then decompose to release damaging reactive oxygen species. We tested whether such redox cycling of alloxan occurs in the human erythrocyte, a cell with stronger antioxidant defenses than beta-cells. Erythrocytes incubated with increasing concentrations of alloxan progressively accumulated dialuric acid, as measured directly by HPLC with electrochemical detection. At concentrations up to 2 mM, alloxan decreased cellular GSH slightly, but did not affect erythrocyte contents of ascorbate or alpha-tocopherol. Intracellular H2O2 generation, measured as inhibition of endogenous catalase activity in the presence of 3-amino-1,2,4-triazole (aminotriazole), was decreased by alloxan. Despite its failure to induce significant oxidant stress in erythrocytes, 2 mM of alloxan doubled the activity of the hexose monophosphate pathway (HMP). This likely reflected consumption of reducing equivalents during reduction of alloxan to dialuric acid. Alloxan pretreatment enhanced the ability of erythrocytes to reduce extracellular ferricyanide while protecting alpha-tocopherol in the cell membrane from oxidation by ferricyanide. Ninhydrin, a hydrophobic derivative of alloxan, showed similar effects, but caused progressive GSH depletion and cell lysis at concentrations above 50 microM. The ability of alloxan to enhance ferricyanide reduction and to spare alpha-tocopherol suggests that dialuric acid or other reducing species within the cells can protect or recycle alpha-tocopherol and donate electrons to a transmembrane transfer process. This behavior resembles that observed for the dehydroascorbate (DHA)/ascorbate pair, and leads to the unexpected conclusion that alloxan increases the reducing capacity of the erythrocyte.
Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232-6303, USA.
J L
JL
Davis
S
S
Mendiratta
J M
JM
May
D000496:Alloxan / Q000378:metabolism*; D001205:Ascorbic Acid / Q000378:metabolism; D003683:Dehydroascorbic Acid / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D005292:Ferricyanides / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D009555:Ninhydrin / Q000378:metabolism; D010084:Oxidation-Reduction; D010427:Pentose Phosphate Pathway; D014810:Vitamin E / Q000378:metabolism
D005292:Ferricyanides; C007931:hexacyanoferrate III; D014810:Vitamin E; D000496:Alloxan; D009555:Ninhydrin; D001205:Ascorbic Acid; D003683:Dehydroascorbic Acid