A chemiluminescence assay detecting the antioxidative effects of glutathione and uric acid on erythrocytes and hemolysates exposed to t-butyl hydroperoxide.
The aim of this study was to compare the antioxidative effects of glutathione (GSH) and uric acid (UA) on erythrocytes and hemolysates exposed to t-butyl hydroperoxide, by using a chemiluminescence (CL) technique. CL formation was induced by t-butyl hydroperoxide in an experimental system consisting of erythrocytes and hemolysates in Tris-HCl buffer (pH 7.4). GSH or UA was added as an antioxidant. In erythrocytes, 10 microM and 50 microM concentrations of either GSH or UA reduced the maximum chemiluminescence values (MCVs) significantly when compared to those of controls. In erythrocytes, MCVs observed in the 50 microM GSH group were significantly lower than those of the 50 microM UA group. In hemolysates, while 10 microM concentration of GSH or UA did not alter MCVs, 50 microM concentration of either GSH or UA reduced MCVs. The reduction of MCVs observed in the 50 microM GSH group was greater than that of the 50 microM UA group. When the effects of equimolar concentrations of GSH or UA on erythrocytes and hemolysates were compared, the MCVs of hemolysates were lower than those of erythrocytes. These results can be attributed to the fact that antioxidant agents react much more easily in the hemolysate medium.
Gümüşlü
S
S
Department of Biochemistry, Faculty of Medicine, Akdeniz University, Arapsuyu, Antalya, Turkey.
Serteser
M
M
Aydin
M
M
Yücel
G
G
D000975:Antioxidants / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects*; D005978:Glutathione / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D010545:Peroxides / Q000009:adverse effects*; D014527:Uric Acid / Q000494:pharmacology*; D020122:tert-Butylhydroperoxide
D000975:Antioxidants; D010545:Peroxides; D014527:Uric Acid; D020122:tert-Butylhydroperoxide; D005978:Glutathione