Öz
In this study, we aimed to investigate the DNA damages and the variations among sperm quality levels in semen samples from healthy individuals and oligozoospermic, insulated asthnenozoospermic, insulated teratozoospermic and oligoasthenoteratozoospermic patients with the factors effecting spermatozoon quality. The sperm samples from healthy individuals and patients diagnosed as insulated oligozoospermia, insulated asthenozoospermia, insulated teratozoospermia, and oligoastheoteratozoospermia in separated groups were used. The number of sperm samples was detected by using Makler camera and their morphologies were evaluated by coloring with Spermac Coloration Assay. To determine the DNA damages among groups, Alkaline Comet Assay method was carried out. According to morphologic results of our study, while the sperm morphology belonging to normozoospermia group was within the normal ranges, the normal sperm morphology in insulated asthenozospermia and insulated oligozoospermia groups was %4 and no significant sperm morphology anomalies were found. In insulated teratozoospermia and oligoasthenoteratozoospermia groups, normal sperm morphology percentages were below %4 and the anomaly percentages were higher than those in the other groups. The results we obtained by using Comet Assay were consistent with morphologic evaluations. As a result, we believe that the Comet Assay can also be guide for determining DNA damages in addition to routine semen analysis and morphologic evaluations.
Anahtar Kelimeler
Kaynakça
- 1. World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th ed. Geneva: World Health Organization 2010: 70- 107.
- 2. Agarwal A, Allamaneni SS. Sperm DNA damage assessment: a test whose time has come. Fertil Steril 2005; 84: 850-853.
- 3. Zini A, Sigman M. Are tests of sperm DNA damage clinically useful? Pros and cons. J Androl 2009; 30: 219-229.
- 4. Zini A, Bielecki R, Phang D, Zenzes MT. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril 2001; 75: 674-677.
- 5. Franken DR, Franken CJ, de la Guerre H, de Villiers A. Normal sperm morphology and chromatin packaging: comparison between aniline blue and chromomysin A3 staining. Andrologia 1999; 31: 361-366.
- 6. Sakkas D, Mariethoz E, Manicardi G et al. Origin of DNA damage in ejaculated human spermatozoa. Reviews of Reproduction 1999; 4: 31-37.
- 7. Ambe AK, Mondragon EC, Gonzales SE. Impact of spermatozoid head anomalies as predictor factor of nondetermined infertility. Ginecol Obstet Mex 2008; 76: 151-155.
- 8. Kruger TF, Ackermann SB, Simmons KF, Swanson RJ, Brugo SS. A quick, reliable staining technique for human sperm morphology. Archives of Andrology 1987; 18: 275-277.
- 9. Arabi M. Bull spermatozoa under mercury stress. Reprod Dom Anim 2005; 40: 454-459.
- 10. Haines G, Marples B, Daniel P, Morris I, DNA damage in human and Mouse spermatozoa after in vitro-irradiation assessed by the comet assay. Adv Exp Med Biol 1998; 444: 789–791.
- 11. Sarıozkan S, Canturk F, Yay A, Akcay A. The effect of different storage temperature on sperm parameters and DNA damage in liquid stored new zealand rabbit spermatozoa. Kafkas Univ Vet Fak Derg 2012; 18: 475–480.
- 12. Verit FF, Verit A, Kocyigit A et al. No increase in sperm DNA damage and seminal oxidative stress in patients with idiopathic infertility. Arch Gynecol Obstet 2006; 274: 339–344.
- 13. Macleod J. Human male infertility. Obstet Gynecol Surv 1971; 26: 335-351.
- 14. Comhaire FH, de Kretser DM, Farley TM, Rowe PJ. Towards more objektivitiy in diagnosis and management of male interfility. Int J Androlo 1987; 7: 1- 53.
- 15. Menkveld R, Lacquet FA, Kruger TF et al. Effects of different staining and washing procedures on the results of human sperm morphology evaluation by manual and computerised medtods. Andrologia 1997; 29: 1-7.
- 16. Bostofte E, Serup J, Rebbe H. Relation between morphologically abnormal spermatozoa and pregnancies obtained during a twenty-year follow-up period. Int J Androl 1982; 5: 379-386.
- 17. Sharbatoghli M, Valojerdi MR, Amanlou M, Khosravi F, Jafar-abadi MA. Relationship of sperm DNA fragmentation, apoptosis and dysfunction of mitochondrial membrane potential with semen parameters and ART outcome after intracytoplasmic sperm injection. Arch Gynecol Obstet 2012; 286:1315-1322.
- 18. Avendano C, Franchi A, Duran H, Oehninger S. DNA fragmentation of normal spermatozoa negatively impacts embryo quality and intracytoplasmic sperm injection outcome. Fertility and Sterility 2010; 94: 549-557.
- 19. Robinson L, Gallos ID, Conner SJ et al. The effect of sperm DNA fragmentation on miscarriage rates: a systematic review and metaanalysis. Human Reproduction 2012; 27: 1-10.
- 20. Zhang L, Wang L, Zhang X et al. Sperm chromatin integrity may predict future fertility for unexplained recurrent spontaneous abortion patients. International Journal of Andrology 2012; 35: 752-757.
- 21. Chi HJ, Chung DY, Choi SY et al. Integrity of human sperm DNA assessed by the neutral comet assay and its relationship to semen parameters and clinical outcomes for the IVF-ET program. Clin Exp Reprod Med 2011; 381: 10-17.
- 22. Saleh R, Agarwal A, Nada E et al. Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Fertility and Sterility 2003; 79: 1597-1603.
- 23. Ribas-Maynou J, Garcia-Peiro A, Fernandez-Encinas A et. al. Comprehensive analysis of sperm DNA fragmentation by five different assays: TUNEL assay, SCSA, SCD test and alkaline and neutral Comet assay. American Society Andrology 2013; 25: 2047-2927.
- 24. Hughes MC, McKelvey-Martin VJ, Lewis SE. Human sperm DNA integrity assessed by the Comet and ELISA assays. Mutagenesis 1999; 14: 71-75.
- 25. Enciso M, Sarasa J, Agarwal A, Fernandez JL, Gosalvez J. A two-tailed Comet assay for assessing DNA damage in spermatozoa. Reproductive BioMedicine 2009; 18: 609-616.
- 26. Ribas-Maynou J, Garcı´a-Peiro A, Abad C et al. Alkaline and neutral Comet assay profiles of sperm DNA damage in clinical groups. Human Reproduction 2012; 27: 652-658.
- 27. Ngo AD, Taylor R, Roberts CL, Nguyen TV. Association between Agent Orange and birth defects: systematic review and meta-analysis. Int J Epidemiol 2006; 355: 1220–1230.
- 28. Host E, Lindenberg S, Kahn J, Christensen F. DNA strand breaks in human sperm cells: a comparison between men with normal and oligozoospermic sperm samples. Acta Obstet Gynecol Scand 1999; 78: 336–339.
- 29. Simon L, Murphy K, Shamsi MB et al. Paternal influence of sperm DNA integrity on early embryonic development. Human Reproduction 2014; 29: 2402-2412.
Öz
Bu çalışmada, izole oligozoospermi, izole astenozoospermi, izole teratozoospermi ve oligoastenoteratozoospermi hastalarından ve sağlıklı bireylerden alınan semen örneklerinde sperm kalite düzeyleri arasındaki farklılıkların ve DNA hasarının incelenmesi amaçlanmıştır. Çalışmada, izole oligozoospermi, izole astenozoospermi, izole teratozoospermi, oligoastenoteratozoospermi tanısı alan hastalar ile sağlıklı erkek bireylerden alınan sperm örnekleri kullanılmıştır. Sperm örneklerinin sayı ve motilitesi Makler kamera ile belirlendi ve Spermac boyama yöntemi ile boyanarak morfolojileri değerlendirildi. Gruplar arasında sperm hücresi DNA hasarını belirlemek amacıyla alkali comet assay yöntemi kullanıldı. Çalışmamızın morfolojik bulgularına göre; normozoospermi grubuna ait sperm morfolojisi normal sınırlar içinde iken, izole astenozoospermi ve izole oligozoospermi gruplarında normal sperm morfolojisi %4 idi ve kayda değer sperm morfoloji anomalilerine rastlanmadı. İzole teratozoospermi ve oligoastenoteratozoospermi gruplarında ise, normal sperm morfoloji yüzdeleri %4’ ün altında olup, anomali yüzdesi diğer gruplara göre daha fazlaydı. Comet ile incelediğimiz sperm DNA hasarı bulguları morfolojik değerlendirmeler ile uyumluydu. Sonuç olarak, rutin semen analizi ve morfolojik değerlendirmelerin yanı sıra DNA hasarını incelemekte comet assay’in de yol gösterici nitelikte kullanılabileceği kanaatindeyiz.
Anahtar Kelimeler
Kaynakça
- 1. World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th ed. Geneva: World Health Organization 2010: 70- 107.
- 2. Agarwal A, Allamaneni SS. Sperm DNA damage assessment: a test whose time has come. Fertil Steril 2005; 84: 850-853.
- 3. Zini A, Sigman M. Are tests of sperm DNA damage clinically useful? Pros and cons. J Androl 2009; 30: 219-229.
- 4. Zini A, Bielecki R, Phang D, Zenzes MT. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril 2001; 75: 674-677.
- 5. Franken DR, Franken CJ, de la Guerre H, de Villiers A. Normal sperm morphology and chromatin packaging: comparison between aniline blue and chromomysin A3 staining. Andrologia 1999; 31: 361-366.
- 6. Sakkas D, Mariethoz E, Manicardi G et al. Origin of DNA damage in ejaculated human spermatozoa. Reviews of Reproduction 1999; 4: 31-37.
- 7. Ambe AK, Mondragon EC, Gonzales SE. Impact of spermatozoid head anomalies as predictor factor of nondetermined infertility. Ginecol Obstet Mex 2008; 76: 151-155.
- 8. Kruger TF, Ackermann SB, Simmons KF, Swanson RJ, Brugo SS. A quick, reliable staining technique for human sperm morphology. Archives of Andrology 1987; 18: 275-277.
- 9. Arabi M. Bull spermatozoa under mercury stress. Reprod Dom Anim 2005; 40: 454-459.
- 10. Haines G, Marples B, Daniel P, Morris I, DNA damage in human and Mouse spermatozoa after in vitro-irradiation assessed by the comet assay. Adv Exp Med Biol 1998; 444: 789–791.
- 11. Sarıozkan S, Canturk F, Yay A, Akcay A. The effect of different storage temperature on sperm parameters and DNA damage in liquid stored new zealand rabbit spermatozoa. Kafkas Univ Vet Fak Derg 2012; 18: 475–480.
- 12. Verit FF, Verit A, Kocyigit A et al. No increase in sperm DNA damage and seminal oxidative stress in patients with idiopathic infertility. Arch Gynecol Obstet 2006; 274: 339–344.
- 13. Macleod J. Human male infertility. Obstet Gynecol Surv 1971; 26: 335-351.
- 14. Comhaire FH, de Kretser DM, Farley TM, Rowe PJ. Towards more objektivitiy in diagnosis and management of male interfility. Int J Androlo 1987; 7: 1- 53.
- 15. Menkveld R, Lacquet FA, Kruger TF et al. Effects of different staining and washing procedures on the results of human sperm morphology evaluation by manual and computerised medtods. Andrologia 1997; 29: 1-7.
- 16. Bostofte E, Serup J, Rebbe H. Relation between morphologically abnormal spermatozoa and pregnancies obtained during a twenty-year follow-up period. Int J Androl 1982; 5: 379-386.
- 17. Sharbatoghli M, Valojerdi MR, Amanlou M, Khosravi F, Jafar-abadi MA. Relationship of sperm DNA fragmentation, apoptosis and dysfunction of mitochondrial membrane potential with semen parameters and ART outcome after intracytoplasmic sperm injection. Arch Gynecol Obstet 2012; 286:1315-1322.
- 18. Avendano C, Franchi A, Duran H, Oehninger S. DNA fragmentation of normal spermatozoa negatively impacts embryo quality and intracytoplasmic sperm injection outcome. Fertility and Sterility 2010; 94: 549-557.
- 19. Robinson L, Gallos ID, Conner SJ et al. The effect of sperm DNA fragmentation on miscarriage rates: a systematic review and metaanalysis. Human Reproduction 2012; 27: 1-10.
- 20. Zhang L, Wang L, Zhang X et al. Sperm chromatin integrity may predict future fertility for unexplained recurrent spontaneous abortion patients. International Journal of Andrology 2012; 35: 752-757.
- 21. Chi HJ, Chung DY, Choi SY et al. Integrity of human sperm DNA assessed by the neutral comet assay and its relationship to semen parameters and clinical outcomes for the IVF-ET program. Clin Exp Reprod Med 2011; 381: 10-17.
- 22. Saleh R, Agarwal A, Nada E et al. Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Fertility and Sterility 2003; 79: 1597-1603.
- 23. Ribas-Maynou J, Garcia-Peiro A, Fernandez-Encinas A et. al. Comprehensive analysis of sperm DNA fragmentation by five different assays: TUNEL assay, SCSA, SCD test and alkaline and neutral Comet assay. American Society Andrology 2013; 25: 2047-2927.
- 24. Hughes MC, McKelvey-Martin VJ, Lewis SE. Human sperm DNA integrity assessed by the Comet and ELISA assays. Mutagenesis 1999; 14: 71-75.
- 25. Enciso M, Sarasa J, Agarwal A, Fernandez JL, Gosalvez J. A two-tailed Comet assay for assessing DNA damage in spermatozoa. Reproductive BioMedicine 2009; 18: 609-616.
- 26. Ribas-Maynou J, Garcı´a-Peiro A, Abad C et al. Alkaline and neutral Comet assay profiles of sperm DNA damage in clinical groups. Human Reproduction 2012; 27: 652-658.
- 27. Ngo AD, Taylor R, Roberts CL, Nguyen TV. Association between Agent Orange and birth defects: systematic review and meta-analysis. Int J Epidemiol 2006; 355: 1220–1230.
- 28. Host E, Lindenberg S, Kahn J, Christensen F. DNA strand breaks in human sperm cells: a comparison between men with normal and oligozoospermic sperm samples. Acta Obstet Gynecol Scand 1999; 78: 336–339.
- 29. Simon L, Murphy K, Shamsi MB et al. Paternal influence of sperm DNA integrity on early embryonic development. Human Reproduction 2014; 29: 2402-2412.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Sağlık Kurumları Yönetimi |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 25 Eylül 2020 |
| Gönderilme Tarihi | 1 Kasım 2019 |
| Yayımlandığı Sayı | Yıl 2020 Cilt: 29 Sayı: 2 |