In vitro effects of high glucose concentrations on membrane protein oxidation, G-actin and deformability of human erythrocytes.
The object of this study was to examine the effect of elevated in vitro glucose concentrations on protein modification and functional changes in human erythrocytes. Groups were exposed to 5-45 mM glucose concentrations. The time effect of any changes was also evaluated. In erythrocyte ghosts, protein glycation and oxidation were evaluated using spectrophotometric methods. G-actin was measured by a DNase I inhibition assay in cell lysates. Erythrocyte deformability was assessed using a cell transit analyser. At 24 h, a significant protein oxidation (at 25 and 45 mM glucose; p < 0.05), and G-actin increase was observed for all concentrations (p < 0.05). At 48 h, a significant increase in glycation (25 and 45 mM glucose; p < 0.05), protein oxidation (p < 0.05), and G-actin (p < 0.05) was observed in all groups. A significant positive correlation was observed between glucose /protein oxidation, glucose/G-actin and protein oxidation/G-actin at 24 and 48 h. Our findings show that the oxidative effect of glucose on erythrocytes depends on concentration and incubation time. We also present the first evidence of increased G-actin in human erythrocytes exposed to high glucose concentrations as a diabetes model.
Resmi
Halil
H
Department of Biochemistry, Dokuz Eylül University Medical School, Izmir, Turkey.
Akhunlar
Hülya
H
Temiz Artmann
Ayşegül
A
Güner
Gül
G
D000199:Actins / Q000378:metabolism*; D002945:Cisplatin / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004907:Erythrocyte Deformability / Q000187:drug effects*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D010084:Oxidation-Reduction / Q000187:drug effects
D000199:Actins; D003432:Cross-Linking Reagents; D005947:Glucose; D002945:Cisplatin