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

Abstract

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.

In Vitro Protective Effect of Phikud Navakot Extraction on Erythrocyte

“…Echinocyte, acanthocyte, and codocyte are erythrocytic membrane defect. Echinocyte formation has been described in H 2 O 2 -induced oxidative stress on red blood cells [24, 25]. In our study, H 2 O 2 -induced oxidative stress caused a number of echinocytes predominately and knizocytes subordinately.…”

“…It can take up more water as compensation for a given amount of osmotic stress. Recently there is an evidence describing that echinocyte formation in H 2 O 2 -induced oxidative stress is caused by SO 4 2− transported defect [24]. Therefore, cellular volume of echinocyte may increase to the highest degree before hemolysis [26].…”

Interaction of Catechins with Human Erythrocytes

“…However, other parameters may be more sensitive to demonstrate the protective effects of lower antioxidant concentrations. One such parameter is the rate of Band 3-mediated sulfate transport, which has been demonstrated to be affected by hydrogen peroxide under conditions inducing neither lipid peroxidation nor thiol oxidation [49].…”

HbE/β-Thalassemia and Oxidative Stress: The Key to Pathophysiological Mechanisms and Novel Therapeutics

“…There is evidence mounting that the inflammatory cascade is initiated by RBC ROS-mediated oxidative stress that directly impacts Band 3 (aka AE1 anion exchanger), an important transmembrane protein in erythrocytes that plays a major role controlling ion transport, possibly NOx transport (71), glycolysis regulation by exchanging glycolytic enzymes with bound Hb or hemichromes (28,101,204), and maintains the linkage between the cytoskeleton and the membrane (54,55,116,131,146). The consequence of oxidative stress appears to originate from excess levels of H 2 O 2 within the RBCs that originate from a progressive loss of the enzymatic detoxification machinery that occurs when RBCs deteriorate due to normal and oxidative damage-accelerated aging (113).…”