PURPOSE: To investigate the extent of γ-irradiation-induced oxidative protein and lipid damage in long-term (up to 21 days) cold stored (4°C) erythrocytes (RBC) and in plasma from whole blood anticoagulated with acid-citrate-dextrose (ACD-A). MATERIALS AND METHODS: Lipid peroxidation, protein carbonyl group (CO) and thiol levels were quantified by the amount of thiobarbituric acid-reactive substances (TBARS), enzyme-linked immunosorbent assay (ELISA) and with Ellman reagent, respectively. RESULTS: Irradiation (40-50 Gy) enhanced lipid peroxidation in the RBC membrane (at day 1 and after 21 storage days); the increase was storage time-dependent. In pre-irradiated (30-50 Gy) and long-term stored RBC membrane protein CO level was higher vs. non-irradiated. Irradiation resulted in RBC membrane protein thiol level elevation, most likely being a result of conformational changes and/or the polypeptide chain fragmentation. Similar to RBC, irradiation of plasma resulted in the increased TBARS generation. In plasma, significant protein CO elevation (at dose of 50 Gy) and protein thiol reduction (30-50 Gy) was observed. CONCLUSION: These findings clearly indicate that irradiation at clinically relevant doses enhances the degree of lipid peroxidation and oxidative protein damage in the membranes of stored RBC. The oxidative stress markers may be considered as additional parameters for RBC quality assessment in the blood banks.
PURPOSE: To investigate the extent of γ-irradiation-induced oxidative protein and lipid damage in long-term (up to 21 days) cold stored (4°C) erythrocytes (RBC) and in plasma from whole blood anticoagulated with acid-citrate-dextrose (ACD-A). MATERIALS AND METHODS:Lipid peroxidation, protein carbonyl group (CO) and thiol levels were quantified by the amount of thiobarbituric acid-reactive substances (TBARS), enzyme-linked immunosorbent assay (ELISA) and with Ellman reagent, respectively. RESULTS: Irradiation (40-50 Gy) enhanced lipid peroxidation in the RBC membrane (at day 1 and after 21 storage days); the increase was storage time-dependent. In pre-irradiated (30-50 Gy) and long-term stored RBC membrane protein CO level was higher vs. non-irradiated. Irradiation resulted in RBC membrane protein thiol level elevation, most likely being a result of conformational changes and/or the polypeptide chain fragmentation. Similar to RBC, irradiation of plasma resulted in the increased TBARS generation. In plasma, significant protein CO elevation (at dose of 50 Gy) and protein thiol reduction (30-50 Gy) was observed. CONCLUSION: These findings clearly indicate that irradiation at clinically relevant doses enhances the degree of lipid peroxidation and oxidative protein damage in the membranes of stored RBC. The oxidative stress markers may be considered as additional parameters for RBC quality assessment in the blood banks.