Study on Oxidative Stress in Thalassemic Red Blood Cells
Keywords:
Oxidative stress, Thalassemia, Flow cytometryAbstract
Abstract: Thalassemia red blood cells (RBCs) are susceptible to oxidative stress, which play a major role in hemolysis. In order to evaluate the oxidative stress in RBCs, we measured the mean fluorescent intensity of dichlorofluorescein (DCF) in RBCs after incubation either with or without 2 mM HO, by flow cytometry. The subjects were 20 healthy adults, 10 alpha-thalassemia or Hemoglobin (Hb) H patients, 12 un-splenectomized and 11 splenectomized B-thal/Hb E patients. For unoxidized
RBCs, significantly higher fluorescent intensity of DCF were observed in thalassemic RBCs, especially in Hb H disease (10.5-7.0), when compared with healthy subjects (2.2+0.4) (p < 0.001), unsplenectomized B-thal/Ho E (40土1.5) (p = 0.008), but no statistical significant difference was found when compared with splenectomized B-thal/Hb E (7.1+3.8) (p > 0.05). Flow cytometric analysis of HO, - oxidized RBCs in all groups showed significantly higher DCF intensity than unoxidized RBCs (p <0.001). The HO, - oxidized RBCs of Hb H disease (300.1+191.2) were higher than unsplenectomized (56.8+50.4) and splenectomized B-thal/Hb E (79.2+51.7) (p < 0.001, 0.005, respectively). There were no significant difference between un-splenectomized and splenectomized B-thal/Hb E (p > 0.05). These findings suggest that there are higher oxidative stress in thalassemic RBCs than healthy subjects and Hb H RBCs seem to have higher oxidative stress than B-tha/Ho E.
Downloads
References
Weatherall DJ, Cleg JB. The thalassemia syndromes. 3rd ed. Oxford, Blackwell, 1981.
Wasi P, Na-Nakom S, Pootrakul S, et al Alpha- and beta-thalassemia in Thailand. Ann NY Acad Sci 1969;165.60-82
Wasi P, Na-Nakorn S, Pootrakul S. The alpha-thalassemias. Clin Haematol 1974;3:383-410.
Wasi P. Hemoglobinopathies in Southeast Asia. In: Bowman JE, ed. Distribution and evaluation and evolution of the hemoglobin gene loci. New York. Elsevier;1983:176-208.
Schrier SL, Mohandas N. Globin-chain specificity of oxidation-induced changes in red blood cell membrane properties. Blood 1992;79:1586-92.
Shinar E, Rachmilewitz EA. Oxidative denaturation of red blood cells in thalassemia. Semin Hematol 1990;27:70-82.
Shinar E, Rachmilewitz EA, Lux SE. Differing erythrocyte membrane skeletal protein defects in alpha- and beta-thalassemia J Clin Invest 1989;83:404-10.
Schrier SL, Rachmilewitz EA, Mohandas N. Cellular and membrane properties of alpha- and beta-thalassemic erythrocytes are different. Blood 1989;74:2194-202.
Hebbel RP, Leung A, Mohandas N. Oxidation-induced changes in microrheological properties of the red blood cell membrane. Blood 1990;76:1015-22.
Yuan J, Bunyaratvej A, Fucharoen S, Fung C, Shinar E, Schrier SL. The instability of the membrane skeleton in thalassemic red blood cells. Blood 1995,86:3945-50.
Grinberg LN, Rachmilewitz EA, Kitrossky N, Chevion M. Hydroxyl radical generation in -thalassemia red blood cells. Free Radic Biol Med 1995;18:611-5.
Prasartkaew S, Bunyaratve] A, Fucharoen S, Wasi P. Oxidative stress and antioxidative enzymes in hemoglobin H disease. Birth Defects 1987;23:193-8.
Yenchitsomanus P, Wasi P. Increased erythrocyte superoxide dismutase activities in Bo-thalassaemia/HbE and in haemoglobin H diseases. J Clin Pathol 1983;36:329-33.
Ong-Ajyooth S, Suthipark K, Shumnumsirivath D, et al. Oxidative stress and antioxidants in B-thalassaemia/Haemoglobin E.J Med Assoc Thai 1987;70:270-4.
Vatanavicharn S, Anuwatanakulchai M, Yenchitsomanus P, Sudhikol C. Relationship of serum vitamin E, erythrocyte nonheme iron, and malonyldialdehyde (lipid membrane peroxidation product) in thalassemia. Birth Defects 1987;23:207-11.
Rachmilewitz EA, Lubin BH, Shohet SB. Lipid peroxidation in beta-thalassemia major. Blood 1976,47:495-505.
Stocks J, Offerman EL, Modell C'B, et al. The susceptibility to autoxidation of human red cell lipids in health and disease. Br J Haematol 1972;23:713-24.
De Zwart LL, Meerman JH, Commandeur JN, Vermeulen NP. Biomarkers of free radical damage applications in experimental animals and in humans. Free Radic Biol Med 1999;26:202-26.
Robinson JP. Oxgen and nitrogen reactive metabolites and phagocytic cells. In: Robinson JP, Babcock GF, eds. Phagocyte function. A guide for research and clinical evaluation. New York: Wiley-Liss, 1998:217-52.
Amer J, Goldfarb A, Fibach E. Flow cytometric measurement of reactive oxygen species production by normal and thalassemic red blood cell. Eur J Haematol 2003;70:84-90.
Polliack A, Rachmilewitz EA. UItrastructural studies in B-thalassemia major. Br J Haematol 1973;24:319-26.
Bauminger ER, Cohen SG, Ofer S, Rachmilewitz EA. Quantitative studies of ferritin like iron in erythrocytes of thalassemia. Sickle-cell anemia, and hemoglobin Hammersmith with Mossbauer spectroscopy. Proc Natl Acad Sci USA 1979;76:939-44.
Jacobs A, Peters SW, Bauminger ER, et al. Ferritin concentration in normal and abnormal erythrocytes measured by immunoradiometric assay with antibodies to heart and spleen ferritin and Mossbauer spectroscopy. Br J Haematol 1981,49:201-7.
Brunori M, Falcioni G, Fioretti E, et al. Formation of superoxide in the autoxidation of the isolated alpha and B-chains of human hemoglobin and its involvement in hemichrome precipitation. Eur J Biochem 1975;53:99-104.
Winterboume CC, McGrath BM, Carrell RW. Reactions involving superoxide and normal and unstable haemoglobins. Biochem J 1976;155:443-502.
Advani R, Sorenson S, Shinar E, Lande W, Rachmilewitz EA, Schrier SL. Characterization and comparison of the red blood cell membrane damage in severe human alpha- and B-thalassemia. Blood 199279.1058-63.
Vigi V, Volpato S, Gaburro D, et al. The correlation between red-cell survival and excess of O-globin synthesis in B-thalassaemia. Br J Haematol 1969;16:25-30.
Polliack A, Yataganas X, Rachmilewitz EA. UItra-structure of the inclusion bodies and nuclear abnormalities B-thalassemic erythroblasts. Ann NY Acad Sci 1974;232:261-82.
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
