Antioxidant Capacity in Seminal Plasma of Transfusion-Dependent β-Thalassemic Patients[*]

Abstract

Background

Homozygous β-thalassemia is a haemolytic disorder with a high potential for oxidative damage, due to the high circulating iron levels. Enzymatic and non enzymatic antioxidant capacities, as well as lipoperoxide content, were investigated in seminal plasma of these patients to evaluate a possible oxidative stress.

Methods

Semen samples from 10 transfusion-dependent β-thalassemic patients and 18 control subjects were examined. The assessment of the seminal antioxidant capacity included spectrophotometrical assays for determination of superoxide dismutase and catalase activity and of the total antioxidant status value. Furthermore, malondialdehyde level was detected as marker of lipoperoxidation.

Results

All the β-thalassemic patients showed high serum ferritin levels, progressive sperm motility below 50 %, and normal sperm count (median: 43 × 10⁶ sperm/ml). Increased superoxide dismutase (p < 0.01) and catalase (p < 0.001) activities, but unaltered total antioxidant status values, were detected in the patients' seminal plasma. Furthermore, augmented malondialdehyde levels (p < 0.001) were measured in the patients.

Conclusions

Seminal antioxidant pattern of iron overloaded β-thalassemic patients indicated the hyperactivation of the enzymatic free-radical scavengers which could be explained as a compensatory mechanism to possible high levels of reactive oxygen species. Furthermore, the increase of seminal lipoperoxidation suggested an oxidative stress in semen of these patients and it could have contributed to the impairment of sperm motility.

Key words

Catalase - human semen - iron overload - superoxide dismutase - thalassemia major - total antioxidant status

1 Financial support: This work was supported by “Ministero dell'Università e della Ricerca Scientifica e Tecnologica”.

References

• 1 Aitken R J, Buckingham D, Carreras A, Irvine D S. Superoxide dismutase in human sperm suspension: relationship with cellular composition, oxidative stress, and sperm function.  Free Radical Med. 1996;  21 495-504
• 2 Alvarez J, Touchstone J C, Blasco L, Storey B T. Spontaneous lipid peroxidation and production of hydrogen peroxide and superoxide in human spermatozoa: Superoxide dismutase as major enzyme protectant against oxygen toxicity.  J Androl. 1987;  8 338-348
• 3 Alvarez J G, Storey B T. Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa.  Mol Hum Reprod. 1995;  42 334-346
• 4 Carpino A, De Sanctis, Siciliano L, Maggiolini M, Vivacqua A, Pinamonti A, Sisci D, Andò S. Epididymal and sex accessory gland secretions in transfusion-dependent β-thalassemic patients: Evidence of an impaired prostatic function.  Exp Clin Endocrinol Diab. 1997;  105 160-165
• 5 Cighetti G, Duca L, Bortone L, Sala S, Nava I, Fiorelli G, Cappellini M D. Oxidative status and malondialdehyde in β-thalassaemia patients.  Eur J Clin Invest. 2002;  32 (Suppl I) 55-60
• 6 De Luca C, Grandinetti M, Maggio F, Lamba M, Passi S. Blood antioxidant status and urinary levels of catecholamine metabolites in beta-thalassemia.  Free Radic Res. 1999;  30 (6) 453-462
• 7 Gavella M, Lipovac V, Vucic M, Rocic B. Superoxide anion scavenging capacity of human seminal plasma.  Int J Androl. 1996;  19 82-90
• 8 Grinberg L N, Rachmilewitz E A, Kitrossky N, Chevion N. Hydroxyl radical generation in beta-thalassemic red blood cells.  Free Radic Bio Med. 1995;  18 611-615
• 9 Griveau J F, Le Lannou D. Reactive oxygen species and human spermatozoa: physiology and pathology.  Int J Androl. 1997;  20 61-69
• 10 Jeulin C, Soufir J C, Weber P, Laval-Martin D, Calvayrac R. Catalase activity in human spermatozoa and seminal plasma.  Gamete Res. 1989;  24 185-196
• 11 Jones R, Mann T, Shering P. Peroxidative breakdown of phospholipids in human spermatozoa, spermicidal properties of fatty acid peroxides and protective action of seminal plasma.  Fertil Steril. 1979;  31 (5) 531-537
• 12 Kobayashi T, Miyazaki T, Natori M, Nozawa S. Protective role of superoxide dismutase in human sperm motility: Superoxide dismutase activity and lipid peroxide in human seminal plasma and spermatozoa.  Hum Reprod. 1991;  6 987-991
• 13 Kushner J P, Porter J P, Olivieri N F. Secondary iron overload.  Hematology. 2001;  47-61
• 14 Livrea M A, Tesoriere L, Pintaudi A M, Calabrese A, Maggio A, Freisleban H J, D'Arpa D, D'Anna R, Bongiorno A. Oxidative stress and antioxidant status in beta-thalassemia major: iron overload and depletion of lipid-soluble antioxidants.  Blood. 1996;  88 (9) 3608-3614
• 15 Miller N J, Rice-Evans C, Davies M J, Gopinathan V, Milner A. A novel method for measuring antioxidant status in premature neonates.  Clin Sci. 1993;  84 407-412
• 16 Ochsendorf F R. Infections in the male genital tract and reactive oxygen species.  Hum Reprod Update. 1999;  5 399-420
• 17 Olivieri N, Nathan D, MacMillan J, Wayne A, Lin P, McGee A, Marin M, Kore G, Cohen A R. Survival in medically treated patients with homozygos beta-thalassemia.  N Engl J Med. 1994;  331 574-578
• 18 Olivieri N F, Brittenham G M. Iron-chelating therapy and the treatment of thalassemia.  Blood. 1997;  89 739-761
• 19 Pearson H, Cohen A, Giardina P J, Kazazian H. The changing profile of homozygous β-thalassemia: demography, ethnicity, and age distribution of current North American patients and changes in two decades.  Pediatrics. 1996;  97 352-356
• 20 Perera D, Pizzey A, Campbell A, Katz M, Porter J, Petrou M, Irvine D S, Chatterjee R. Sperm DNA damage in potentially fertile homozygous β-thalassaemia patients with iron overload.  Hum Reprod. 2002;  17 (7) 1820-1825
• 21 Pick E, Mizel D. Rapid microassays for the measurement of superoxide and hydrogen peroxide production by macrophages in culture using an automatic enzyme immunoassay reader.  J Immunol Methods. 1981;  46 211-226
• 22 Psihogios V, Rodda C, Reid E, Clark M, Clarke C, Bowden D. Reproductive health in individuals with homozygous β-thalassemia: knowledge, attitudes, and behavior.  Fertil Steril. 2002;  77 (1) 119-127
• 23 Rebulla P. Bood transfusion in beta thalassemia major.  Transfus Med. 1995;  5 247-258
• 24 Reubinoff B E, Har-EI R, Kitrossky N, Friedler S, Levi R, Lewin A, Chevion M. Increased levels of redox-active iron in follicular fluid: a possible cause of free radical-mediated infertility in beta-thalassemia major.  Am J Obstet Gynecol. 1996;  174 914-918
• 25 Saleh R, Agarwal A. Oxidative stress and male infertility: from research bench to clinical practice.  J Androl. 2002;  23 (6) 737-752
• 26 Sharma R, Agarwal A. Role of reactive oxygen species in male infertility.  Urology. 1996;  48 835-850
• 27 Siciliano L, Tarantino P, Longobardi F, Rago V, De Stefano C, Carpino A. Impaired seminal antioxidant capacity in human semen with hyperviscosity or oligoasthenozoospermia.  J Androl. 2001;  22 798-803
• 28 Sikka S C. Oxidative stress and role of antioxidants in normal and abnormal sperm function.  Front Biosci. 1996;  1 e78-86
• 29 Upreti G C, Jensen K, Oliver J E, Duganzich D M, Munday R, and Smith J F. Motility of ram spermatozoa during storage in a chemically defined diluent containing antioxidants.  Anim Reprod Sci. 1997;  48 269-278
• 30 World Health Organization .Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction. Cambridge, UK; Cambridge University Press 1999
• 31 Zurlo M G, De Stefano P, Borgna-Pignatti C, Di Palma A, Piga A, Melevendi C, Di Gregorio F, Burattini M G, Terzoli S. Survival and causes of death in thalassemia major.  Lancet. 1989;  2 27-30

1 Financial support: This work was supported by “Ministero dell'Università e della Ricerca Scientifica e Tecnologica”.