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Journal of Assisted Reproduction and Genetics

, Volume 34, Issue 3, pp 373–381 | Cite as

Effects of reduced seminal enzymatic antioxidants on sperm DNA fragmentation and semen quality of Tunisian infertile men

  • Fatma AtigEmail author
  • Abdelhamid Kerkeni
  • Ali Saad
  • Mounir Ajina
Gamete Biology

Abstract

Purpose

To evaluate levels of sperm DNA fragmentation and enzymatic antioxidant status in seminal plasma of Tunisian fertile and infertile men in order to assess the effects of seminal oxidative stress on sperm DNA integrity and semen quality.

Methods

Semen samples from 100 infertile patients (40 oligoasthenoteratozoospermics, 31 teratozoospermics and 29 asthenozoospermics) and 50 fertile men (controls) were analyzed for DNA fragmentation by TUNEL assay and biochemical parameters. Seminal antioxidant activities (Superoxide dismutase, Glutathione peroxidase and Catalase) and malondialdehyde concentrations were measured spectrophotometrically.

Results

Sperm DNA fragmentation and malondialdehyde levels in infertile groups were more elevated than controls. Nevertheless, the activities of the antioxidant enzymes were significantly lower in abnormal groups compared to normozoospermics. Sperm DNA fragmentation was closely and positively correlated to malondialdehyde levels (r = 0.37, P = 0.008); meanwhile, reduced seminal antioxidant profile was negatively associated to sperm DNA fragmentation. Interestingly, we noted also that sperm DNA fragmentation was negatively correlated to sperm motility (r = −0.54, P < 0.001) and positively associated to the abnormal sperm morphology (r = 0.57, P = 0.002).

Conclusions

This report revealed that increased sperm DNA fragmentation can be due to the impaired seminal enzymatic antioxidant profile and increased Lipid peroxidation. Our results sustain that the evaluation of sperm DNA fragmentation and seminal oxidative biomarkers in infertile men is recommended as a consistent prognostic tool for male infertility assessment.

Keywords

Male infertility Sperm DNA fragmentation Oxidative stress Reactive oxygen species Seminal plasma Antioxidants 

Notes

Acknowledgments

The authors are especially grateful to the study participants. We would like to thank the staff of the “department of Cytogenetic and Reproductive Biology, Soussa, Farhat Hached Hospital” for their help in semen selection and technician assistance.

Competing interests

The authors have declared that no competing interest exists.

References

  1. 1.
    Agarwal A, Saleh RA, Bedaiwy MA. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril. 2003;79:829–43.CrossRefPubMedGoogle Scholar
  2. 2.
    Agarwal A, Sekhon LH. Oxidative stress and antioxidants for idiopathic oligoasthenoteratospermia: Is it justified? Indian J Urol. 2011;27:74–85.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Aitken RJ, De Iuliis GN, McLachlan RI. Biological and clinical significance of DNA damage in the male germ line. Int J Androl. 2009;32:46–56.CrossRefPubMedGoogle Scholar
  4. 4.
    Aitken RJ, De Iuliis GN, Finnie JM, et al. Analysis of the relationships between oxidative stress, DNA damage and sperm vitality in a patient population: development of diagnostic criteria. Hum Reprod. 2010;25:2415–26.CrossRefPubMedGoogle Scholar
  5. 5.
    Alvarez JG, Storey BT. Spontaneous lipid peroxidation in rabbit epididymal spermatozoa: its effect on sperm motility. Biol Reprod. 1982;27:1102–8.CrossRefPubMedGoogle Scholar
  6. 6.
    Atig F, Raffa M, Habib BA, et al. Impact of seminal trace element and glutathione levels on semen quality of Tunisian infertile men. BMC Urol. 2012;12:6.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Atig F, Raffa M, Ali HB, et al. Altered antioxidant status and increased lipid per-oxidation in seminal plasma of tunisian infertile men. Int J Biol Sci. 2012;8:139–49.CrossRefPubMedGoogle Scholar
  8. 8.
    Aziz N, Saleh RA, Sharma RK, et al. Novel association between sperm reactive oxygen species production, sperm morphological defects, and the sperm deformity index. Fertil Steril. 2004;81:349–54.CrossRefPubMedGoogle Scholar
  9. 9.
    Beer B, Sizer W. A spectrophotometry method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952;195:133–9.Google Scholar
  10. 10.
    Brahem S, Mehdi M, Elghezal H, et al. Detection of DNA fragmentation and meiotic segregation in human with isolated teratozoospermia. J Assist Reprod Genet. 2011;28:41–8.CrossRefPubMedGoogle Scholar
  11. 11.
    Brahem S, Mehdi M, ElGhezal H, et al. The effects of male aging on semen quality, sperm DNA fragmentation and chromosomal abnormalities in an infertile population. J Assist Reprod Genet. 2011;28:425–32.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Chen Z, Hauser R, Trbovich A, et al. The relation between human semen characteristics and sperm apoptosis: a pilot study. J Androl. 2006;27:112–20.CrossRefPubMedGoogle Scholar
  13. 13.
    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;38:10–7.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Cohen-Bacrie P, Belloc S, Ménézo YJ, et al. Correlation between DNA damage and sperm parameters: a prospective study of 1,633 patients. Fertil Steril. 2009;91:1801–5.CrossRefPubMedGoogle Scholar
  15. 15.
    Dakouane M, Albert M, Bergere M, et al. Aging and spermatogenesis: an histologic, cytogenetic and apoptosis study. Gynecol Obstet Fertil. 2005;33:659–64.CrossRefPubMedGoogle Scholar
  16. 16.
    David G. Editorial: sperm banks in France. Arch Fr Pediatr. 1975;32:401–4.PubMedGoogle Scholar
  17. 17.
    Erenpreiss J, Spano M, Erenpreisa J, et al. Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl. 2006;8:11–29.CrossRefPubMedGoogle Scholar
  18. 18.
    Giannattasio A, De Rosa M, Smeraglia R, et al. Glutathione peroxidase (GPX) activity in seminal plasma of healthy and infertile males. J Endocrinol Invest. 2002;25:983–6.CrossRefPubMedGoogle Scholar
  19. 19.
    Gornall AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem. 1949;177:751–66.PubMedGoogle Scholar
  20. 20.
    Günzler WA, Kremers H, Flohé L. An improved coupled test procedure for glutathione peroxidase in blood. Z Klin Chem Klin Biochem. 1974;12:444–8.PubMedGoogle Scholar
  21. 21.
    Hassen M, Kurinczuk JJ, Browser C, et al. The risk of major birth defects after intra-cytoplasmic sperm injection and in vitro fertilization. N Engl J Med. 2002;346:725–30.CrossRefGoogle Scholar
  22. 22.
    Huang CC, Lin DP, Tsao HM, et al. Sperm DNA fragmentation negatively correlates with velocity and fertilization rates but might not affect pregnancy rates. Fertil Steril. 2005;84:130–40.CrossRefPubMedGoogle Scholar
  23. 23.
    Markund S, Markund G. Involvement of the superoxide anion radical in autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem. 1974;47:469–74.CrossRefGoogle Scholar
  24. 24.
    Marzec-Wróblewska U, Kamiński P, et al. Zinc and iron concentration and SOD activity in human semen and seminal plasma. Biol Trace Elem Res. 2010;143:167–77.CrossRefPubMedGoogle Scholar
  25. 25.
    Mehdi M, Khantouche L, Ajina M, et al. Detection of DNA fragmentation in human spermatozoa: correlation with semen parameters. Andrologia. 2009;41:383–6.CrossRefPubMedGoogle Scholar
  26. 26.
    Nasr-Esfahani MH, Salehi M, Razavi S, et al. Effect of sperm DNA damage and sperm protamine deficiency on fertilization and embryo development post-ICSI. Reprod Biomed Online. 2005;11:198–205.CrossRefPubMedGoogle Scholar
  27. 27.
    Potts RJ, Notarianni LJ, Jefferies TM. Seminal plasma reduces exogenous oxidative damage to human sperm, determined by the measurement of DNA strand breaks and lipid peroxidation. Mutat Res. 2000;447:249–56.CrossRefPubMedGoogle Scholar
  28. 28.
    Powell SR. The antioxidant properties of zinc. J Nutr. 2000;130(5S Suppl):1447S–54S.PubMedGoogle Scholar
  29. 29.
    Sakkas D, Mariethoz E, Manicardi G, et al. Origin of DNA damage in ejaculated human spermatozoa. Rev Reprod. 1999;4:31–7.CrossRefPubMedGoogle Scholar
  30. 30.
    Sakkas D, Tomlinson M. Assessment of sperm competence. Semin Reprod Med. 2000;18:133–9.CrossRefPubMedGoogle Scholar
  31. 31.
    Saleh RA, Agarwal A, Nelson DR, et al. Increased sperm nuclear DNA damage in normozoospermic infertile men: a prospective study. Fertil Steril. 2002;78:313–8.CrossRefPubMedGoogle Scholar
  32. 32.
    Saleh RA, Agarwal A, Nada EA, et al. Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Fertil Steril. 2003;79:1597–605.CrossRefPubMedGoogle Scholar
  33. 33.
    Shamsi MB, Kumar R, Bahtt A, et al. Mitochondrial DNA mutations in etiopathogenesis of male infertility. Indian J Urol. 2008;24:150–4.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Shamsi MB, Venkatesh S, Tanwar M, et al. DNA integrity and semen quality in men with low seminal antioxidant levels. Mutat Res. 2009;665:29–36.CrossRefPubMedGoogle Scholar
  35. 35.
    Singh NP, Muller CH, Berger RE. Effects of age on DNA doublestrand breaks and apoptosis in human sperm. Fertil Steril. 2003;80:1420–30.CrossRefPubMedGoogle Scholar
  36. 36.
    Sills ES, Fryman JT, Perloe M, et al. Chromatin fluorescence characteristics and standard semen analysis parameters: correlation observed in andrology testing among 136 males referred for infertility evaluation. J Obstet Gynaecol. 2004;24:74–7.CrossRefPubMedGoogle Scholar
  37. 37.
    World Health Organization. WHO laboratory manual for the examination of human semen and semen-cervical mucus interaction. 4 editions. Cambridge, UK: Published on behalf of the World Health Organization by Cambridge University Press; 1999. 1–86.Google Scholar
  38. 38.
    Yagi K. A simple fluorometric assay for lipoperoxide in blood plasma. Biochem Med. 1976;15:212–6.CrossRefPubMedGoogle Scholar
  39. 39.
    Zini A, Fischer MA, Sharir S, et al. Prevalence of abnormal sperm DNA denaturation in fertile and infertile men. Urology. 2002;60:1069–72.CrossRefPubMedGoogle Scholar
  40. 40.
    Zribi N, Chakroun NF, Elleuch H, et al. Sperm DNA fragmentation and oxidation are independent of malondialdehyde. Reprod Biol Endocrinol. 2011;9:47.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Fatma Atig
    • 1
    • 2
    • 3
    Email author
  • Abdelhamid Kerkeni
    • 3
  • Ali Saad
    • 1
    • 2
  • Mounir Ajina
    • 1
    • 2
  1. 1.Unit of Reproductive MedicineUniversity Farhat Hached HospitalSoussaTunisia
  2. 2.Department of Cytogenetic and Reproductive BiologyUniversity Farhat Hached HospitalSoussaTunisia
  3. 3.Research Laboratory of “Trace elements, free radicals and antioxidants”, Biophysical Department, Faculty of MedicineUniversity of MonastirMonastirTunisia

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