Estresse oxidativo em membranas de eritrócito avaliado por ressonância paramagnética eletrônica
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2010-03-26
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Universidade Federal de Goiás
Resumo
The oxidative stress effects promoted by hydrogen peroxide (H2O2) and 2,2 -Azobis(2- methylpropionamidine)dihydrochloride (AAPH) in proteins and lipids of the erythrocyte membrane were investigated by testing the oxidative hemolysis, the formation of malondialdehyde
(MDA) in addition to spectroscopic electron paramagnetic resonance (EPR) of lipid spin label 5-DOXIL stearic acid (5-DSA) and 3-maleimide proxyl (5-MSL) that binds covalently to
erythrocyte s membrane proteins. The spectral parameter 2Ak obtained directly from the EPR spectra of spin label 5-DSA structured in the lipid bilayer of the erythrocyte membrane was sensitive to changes in the dynamics of lipids resulting on the oxidation of membrane proteins. The oxidation of proteins observed for very low concentrations of H2O2 (starting at 100 μM) were confirmed with spin label 5-MSL. Lipid peroxidation indicated the oxidative hemolysis and formation of MDA occurred at concentrations of H2O2 about 8 times larger (starting at 800 μM). Ascorbic acid and -tocopherol protect the membrane by hemolysis and MDA tests, but did not prevent the stiffening of the erythrocyte membrane. The spectra of the spin label 5-MSL revealed the existence of two distinct thiol groups in erythrocyte membrane proteins that differ in their structural configurations and sensitivity to oxidative attack. The SH site that has higher exposure to solvent and less reactivity to spin label 5-MSL was the most vulnerable to oxidation. It is well known that upon oxidation hemoglobin binds to the membrane and the results of this study suggest that this effect may be accompanied by a further increase in the parameter 2Ak of the spin label 5-DSA. Catalase present in erythrocytes proved to be an effective protector of lipid peroxidation and oxidation of membrane proteins induced by hydrogen peroxide.
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MENDANHA NETO, Sebastião Antônio. Estresse oxidativo em membranas de eritrócito avaliado por ressonância paramagnética eletrônica. 2010. 67 f. Dissertação (Mestrado em Fisica) - Universidade Federal de Goiás, Goiânia, 2010.