H2O2-Induced Oxidative Stress Affects SO4= Transport in Human Erythrocytes

• Published in: PloS one 2016, Vol.11 (1), p.e0146485-e0146485
• Main Authors: Morabito, Rossana, Romano, Orazio, La Spada, Giuseppa, Marino, Angela
• Format: Article
• Language: English
• Subjects: Alcoholic beverages; Alcoholism; Antioxidants; Antioxidants - pharmacology; Biological Transport; Blood; Cell Membrane Permeability; Cell Shape; Chlorides; Contrast agents; Contrast media; Curcumin; Curcumin - pharmacology; Cytoskeleton; Deformability; Deformation; Drugs; Environmental science; Erythrocyte Membrane - drug effects; Erythrocyte Membrane - ultrastructure; Erythrocytes; Erythrocytes - drug effects; Erythrocytes - metabolism; Erythrocytes - ultrastructure; Exposure; Flavonoids; Formability; Ghosts; Glutathione; Glutathione - pharmacology; Hemoglobin; Humans; Hydrogen peroxide; Hydrogen Peroxide - pharmacology; Kinetics; Lipids; Malondialdehyde; Malondialdehyde - metabolism; Neurodegenerative diseases; Neurological diseases; Oxidative Stress; Proteins; Reduction; Sulfates - metabolism; Transport
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0146485

The aim of the present investigation was to verify the effect of H2O2-induced oxidative stress on SO4= uptake through Band 3 protein, responsible for Cl-/HCO3- as well as for cell membrane deformability, due to its cross link with cytoskeletal proteins. The role of cytoplasmic proteins binding to Band 3 protein has been also considered by assaying H2O2 effects on hemoglobin-free resealed ghosts of erythrocytes. Oxidative conditions were induced by 30 min exposure of human erythrocytes to different H2O2 concentrations (10 to 300 μM), with or without GSH (glutathione, 2 mM) or curcumin (10 μM), compounds with proved antioxidant properties. Since SO4= influx through Band 3 protein is slower and better controllable than Cl- or HCO3- exchange, the rate constant for SO4= uptake was measured to prove anion transport efficiency, while MDA (malondialdehyde) levels and -SH groups were estimated to quantify the effect of oxidative stress. H2O2 induced a significant decrease in rate constant for SO4= uptake at both 100 and 300 μM H2O2. This reduction, observed in erythrocytes but not in resealed ghosts and associated to increase in neither MDA levels nor in -SH groups, was impaired by both curcumin and GSH, whereas only curcumin effectively restored H2O2-induced changes in erythrocytes shape. Our results show that: i) 30 min exposure to 300 μM H2O2 reduced SO4= uptake in human erythrocytes; ii) oxidative damage was revealed by the reduction in rate constant for SO4= uptake, but not by MDA or -SH groups levels; iii) the damage was produced via cytoplasmic components which cross link with Band 3 protein; iv) the natural antioxidant curcumin may be useful in protecting erythrocytes from oxidative injury; v) SO4= uptake through Band 3 protein may be reasonably suggested as a tool to monitor erythrocytes function under oxidative conditions possibly deriving from alcohol consumption, use of drugs, radiographic contrast media administration, hyperglicemia or neurodegenerative diseases.