HYDROXYL RADICAL GENERATION IN BETA-THALASSEMIC RED-BLOOD-CELLS

To provide more experimental evidence for the proposed role of oxygen free radicals in red blood cell (RBC) damage in beta-thalassemia, hydr oxyl radical generation was studied in thalassemic (Th) vs. normal (N) RBC. (OH)-O-. fluxes were quantified by the conversion of salicylic a cid (SA) into its hydroxylated products, 2,3- and 2,5-dihydroxybenzoic acids (DHBA) and catechol, assayed with HPLC coupled to electrochemic al detection. No significant difference in spontaneous (OH)-O-. genera tion between N-RBC and Th-RBC was found. Ascorbic acid (0.5-3.0 mM) in duced many-fold increases in SA hydroxylation in a dose-dependant mann er in both types of cells. In the presence of ascorbate (1.0 mM), the SA hydroxylated products were determined in Th-RBC vs. N-RBC as follow s (nmol/ml): 2,5-DHBA, 1.45 +/- 0.06 vs. 1.81 +/- 0.05 (p = 0.001); 2, 3-DHBA, 1.89 +/- 0.21 vs. 1.15 +/- 0.08 (p =0.008) and catechol, 0.87 +/- 0.13 vs. 0.38 +/- 0.05 (p =0.006). The results showed significant increase in the total SA hydroxylation in Th-RBC as compared to N-RBC with a tendency to form 2,3-DHBA and catechol at the expanse of 2,5-DH BA. The excessive (OH)-O-. generation in Th-RBC is attributed to the a bnormally high content of redox active iron in the cytosolic and/or me mbrane compartments of these cells.