The relationship between Sperm DNA fragmentation and differential expression of human sperm pro-apoptotic miR-15a/16 and anti-apoptotic BCL-2 gene

Document Type : Original Article

Authors

1 M.Sc. of Anatomy, Cellular and Molecular Biology Research Center, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Assistant Professor, Department of Embryology, Mam Research Center, Tehran, Iran.

3 Associate Professor, Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran, Associate Professor, Department of Biotechnology, School of Advanced Technologies, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

Introduction: Apoptosis is one of the etiologies of sperm DNA damage. Considering the crucial role of micro RNAs in multiple pathophysiological processes, the possibility of controlling the apoptosis process in sperm DNA by micro-RNAs has been proposed. This study was performed with aim to investigate the relationship between sperm DNA fragmentation and expression level of miR-15a/16 and Anti-apoptotic BCL-2 gene.
Methods: This experimental study was performed on 30 patients referred to Tehran Taleghani Hospital infertility center in 2017. After 2-3 days of abstinence, semen was collected and analyzed based on WHO criteria. Sperm preparation was performed using density gradient centrifugation technique. To assess sperm DNA fragmentation, Sperm Chromatin Structure Assay was carried out and based on the level of DNA fragmentation, the samples were divided into two groups: DFI ≥30% and DFI<30%. The expression level of miR-15a/16 and BCL-2 was measured by quantitative Real Time -PCR. Data were analyzed using SPSS software (version 22) and independent t-test. The gene expression level was analyzed using REST 2009 software. P < 0.05 was considered statistically significant.
Results: A significantnegative relationship was found between DFI ≥30% with motility and normal morphology of spermatozoa (p<0.05). The expression level of miR-15a/16 was significantly increased, and the expression level of BCL-2 gene was significantly decreased in patients with DFI ≥30% compared with DFI<30% (p<0.05).
Conclusion: Decreased expression of BCL2 following increased miR-16-1 leads to increased sperm DNA fragmentation. Assessment of miR-15a and apoptosis-related gene can be proposed as a diagnosing tool to ensure of sperm DNA integrity in male factor infertility cases in future.

Keywords

References

  1. Hull MG, Glazener CM, Kelly NJ, Conway DI, Foster PA, Hinton RA, et al. Population study of causes, treatment, and outcome of infertility. Br Med J (Clin Res Ed) 1985; 291(6510):1693-7.
  2. Brugh VM 3rd, Lipshultz LI. Male factor infertility: evaluation and management. Med Clin North Am 2004; 88(2):367-85.
  3. World Health Organization. WHO laboratory manual for the examination and processing of human semen. Geneva: World Health Organization; 2010.
  4. Evgeni E, Charalabopoulos K, Asimakopoulos B. Human sperm DNA fragmentation and its correlation with conventional semen parameters. J Reprod Infertil 2014; 15(1):2-14.
  5. Boushaba S, Belaaloui G. Sperm DNA fragmentation and standard semen parameters in algerian infertile male partners. World J Mens Health 2015; 33(1):1-7.
  6. Osadchuk LV, Tataru DA, Kuznetsova NN, Kleshev MA, Markova EV, Svetlakov AV. Sperm DNA fragmentation: association with semen parameters in young men. Urologiia 2016; 6:118-23.
  7. Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl 2002; 23(1):25-43.
  8. 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(4):674-7.
  9. Avendano C, Franchi A, Duran H, Oehninger S. DNA fragmentation of normal spermatozoa negatively impacts embryo quality and intracytoplasmic sperm injection outcome. Fertil Steril 2010; 94(2):549-57.
  10. Schulte RT, Ohl DA, Sigman M, Smith GD. Sperm DNA damage in male infertility: etiologies, assays, and outcomes. J Assist Reprod Genet 2010; 27(1):3-12.
  11. Lin MH, Kuo-Kuang Lee R, Li SH, Lu CH, Sun FJ, Hwu YM. Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertil Steril 2008; 90(2):352-9.
  12. Choi HY, Kim SK, Kim SH, Choi YM, Jee BC. Impact of sperm DNA fragmentation on clinical in vitro fertilization outcomes. Clin Exp Reprod Med 2017; 44(4):224-31.
  13. Centola GM, Ginsburg KA. Evaluation and treatment of the infertile male. Cambridge: Cambridge University Press; 2004.
  14. Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 1980; 284(5756):555-6.
  15. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35(4):495-516.
  16. Wyllie AH. The genetic regulation of apoptosis. Curr Opin Genet Dev 1995; 5(1):97-104.
  17. O'Brien J, Hayder H, Zayed Y, Peng C. Overview of MicroRNA biogenesis, mechanisms of actions, and circulation. Front Endocrinol 2018; 9:402.
  18. Subramanian S, Steer CJ. MicroRNAs as gatekeepers of apoptosis. J Cell Physiol 2010; 223(2):289-98.
  19. Cimmino A, Calin GA, Fabbri M, Iorio MV, Ferracin M, Shimizu M, et al. miR-15 and miR-16 induce apoptosis by targeting BCL2. Proc Natl Acad Sci U S A 2005; 102(39):13944-9.
  20. Salehi M, Afarinesh MR, Haghpanah T, Ghaffari Novin M, Farifteh F. Impact of sperm DNA fragmentation on ICSI outcome and incidence of apoptosis of human pre-implantation embryos obtained from in vitro matured MII oocytes. Biochem Biophys Res Commun 2019; 510(1):110-5.
  21. Zini A, Boman JM, Belzile E, Ciampi A. Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis. Hum Reprod 2008; 23(12):2663-8.
  22. Zini A, Libman J. Sperm DNA damage: importance in the era of assisted reproduction. Curr Opin Urol 2006; 16(6):428-34.
  23. Virro MR, Larson-Cook KL, Evenson DP. Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil Steril 2004; 81(5):1289-95.
  24. Yamamoto CM, Hikim AP, Lue Y, Portugal AM, Guo TB, Hsu SY, et al. Impairment of spermatogenesis in transgenic mice with selective overexpression of Bcl-2 in the somatic cells of the testis. J Androl 2001; 22(6):981-91.
  25. Mostafa T, Rashed L, Nabil N, Amin R. Seminal BAX and BCL2 gene and protein expressions in infertile men with varicocele. Urology 2014; 84(3):590-5.
  26. Hayashi K, Chuva de Sousa Lopes SM, Kaneda M, Tang F, Hajkova P, Lao K, et al. MicroRNA biogenesis is required for mouse primordial germ cell development and spermatogenesis. PLoS One 2008; 3(3):e1738.
  27. Kotaja N, Lin H, Parvinen M, Sassone-Corsi P. Interplay of PIWI/Argonaute protein MIWI and kinesin KIF17b in chromatoid bodies of male germ cells. J Cell Sci 2006; 119(Pt 13):2819-25.
  28. Khazaie Y, Nasr Esfahani MH. MicroRNA and male infertility: a potential for diagnosis. Int J Fertil Steril 2014; 8(2):113-8.
  29. Abhari A, Zarghami N, Farzadi L, Nouri M, Shahnazi V. Altered of microRNA expression level in oligospermic patients. Iran J Reprod Med 2014; 12(10):681-6.
  30. Abu-Halima M, Hammadeh M, Schmitt J, Leidinger P, Keller A, Meese E, et al. Altered microRNA expression profiles of human spermatozoa in patients with different spermatogenic impairments. Fertil Steril 2013; 99(5):1249-55.
  31. Shaha C, Tripathi R, Mishra DP. Male germ cell apoptosis: regulation and biology. Philos Trans R Soc Lond B Biol Sci 2010; 365(1546):1501-15.
  32. Lian J, Zhang X, Tian H, Liang N, Wang Y, Liang C, et al. Altered microRNA expression in patients with non-obstructive azoospermia. Reprod Biol Endocrinol 2009; 7(1):13.
  33. Curry E, Safranski TJ, Pratt SL. Differential expression of porcine sperm microRNAs and their association with sperm morphology and motility. Theriogenology 2011; 76(8):1532-9.
  34. Wang C, Yang C, Chen X, Yao B, Yang C, Zhu C, et al. Altered profile of seminal plasma microRNAs in the molecular diagnosis of male infertility. Clin Chem 2011; 57(12):1722-31.

The Iranian Journal of Obstetrics, Gynecology and Infertility
Volume 22, Issue 10
December 2019
Pages 42-48

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