Evaluation of the effect of antioxidant resveratrol on sperm biochemical matches and expression of caspase 3 and HSP70 genes in infertile stenotratospermia under freezing conditions

Document Type : Original article

Authors

1 Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Biology, Faculty of Basic Sciences, Kharazmi University, Tehran, Iran

3 Department of Biology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran

Abstract

Introduction: One of the most important measures to increase sperm quality and consequently the success of assisted reproductive technology (ART) is to protect sperm against oxidative reactions during freezing. Zoratrol is an antioxidant compound that protects the body against chronic diseases by scavenging free radicals. The aim of this study was to evaluate the effect of resveratrol on human sperm functional parameters and to evaluate apoptosis and expression of HSP70 and Caspase3 genes after cryopreservation.
Materials and Methods: In this study, 40 men referred to the ACECR Infertility Center in Qom, all of whom met the inclusion criteria, were used as the target population. The target population was divided into two groups of men with normal WHO criteria and a second group of patients with asthenotracheal sperm. After sperm sampling, the sperm was frozen with different concentrations of resveratrol. Semen samples were examined for sperm parameters including concentration, morphology, motility, viability, reactive oxygen species (ROS), apoptosis and expression of HSP70 and caspase 3 genes.
 Results: Resveratrol significantly increased sperm motility by 0.02 and 0.05 mM (P≤0.001), but decreased compared to the control group with resveratrol by 0.25 and 0.1 mM (P≤0.001). With the exception of resveratrol 0.25 mM, all doses of resveratrol, especially 0.05 mM, increased sperm count (P˂0.05). All doses of resveratrol, especially 0.02 mM and 0.05 mM, reduced apoptosis and expression of HSP70 and caspase3 genes.
Conclusion: In general, resveratrol supplementation before cryopreservation significantly increased sperm quality and reduced the side effects of cryopreservation shock.
 

Keywords


  1. Sherman JK. Synopsis of the use of frozen human semen since 1964: state of the art of human semen banking. Fertility and Sterility. 1973 May;24(5):397. https://doi.org/10.1016/s0015-0282(16)39678-9.
  2. Stanic P, Tandara M, Sonicki Z, Simunic V, Radakovic B, Suchanek Comparison of protective media and freezing techniques for cryopreservation of human semen. European Journal of Obstetrics & Gynecology and Reproductive Biology. 2000 Jul 1;91(1):65-70. https://doi.org/10.1016/s0301-2115(99)00255-9.
  3. Samali A, Orrenius S. Heat shock proteins: regulators of stress response and apoptosis. Cell stress & chaperones. 1998 Dec;3(4):228. https://doi.org/10.1379/1466-1268(1998)003.
  4. Whitley D, Goldberg SP, Jordan WD. Heat shock proteins: a review of the molecular chaperones. Journal of Vascular Surgery. 1999 Apr 1;29(4):748-51. https://doi.org/10.1016/s0741-5214(99)70329-0.
  5. Mayer MP, Bukau B. Hsp70 chaperones: cellular functions and molecular mechanism. Cellular and molecular life sciences. 2005 Mar 1;62(6):670. https://doi.org/10.1007/s00018-004-4464-6.
  6. Bukau B, Horwich AL. The Hsp70 and Hsp60 chaperone machines. 1998 Feb 6;92(3):351-66.https://doi.org/10.1016/s0092- 8674(00)80928-9.
  7. Simon MM, Reikerstorfer A, Schwarz A, Krone C, Luger TA, Jäättelä M, Schwarz T. Heat shock protein 70 overexpression affects the response to ultraviolet light in murine fibroblasts. Evidence for increased cell viability and suppression of cytokine release. The Journal of clinical investigation. 1995 Mar 1;95(3):926-33. https://doi.org/10.1172/jci117800.
  8. Sun Y, Chen XY, Liu J, Cheng XX, Wang XW, Kong QY, Li H. Differential caspase-3 expression in noncancerous, premalignant and cancer tissues of stomach and its clinical implication. Cancer Detection and Prevention. 2006 Jan 1;30(2):168-73. https://doi.org/10.1016/j.cdp.2006.02.004.
  9. Kim PK, Armstrong M, Liu Y, Yan P, Bucher B, Zuckerbraun BS, Gambotto A, Billiar TR, Yim JH. IRF-1 expression induces apoptosis and inhibits tumor growth in mouse mammary cancer cells in vitro and in vivo. Oncogene. 2004 Feb;23(5):1125-35. https://doi.org/10.1038/sj.onc.1207023
  10. Guo J, Bourre L, Soden DM, O'Sullivan GC, O'Driscoll C. Can nonviral technologies knockdown the barriers to siRNA delivery and achieve the next generation of cancer therapeutics?. Biotechnology 2011 Jul 1;29(4):402-17. https://doi.org/10.1016/j.biotechadv.2011.03.003.
  11. Khori V, Shalamzari SA, Isanejad A, Alizadeh AM, Alizadeh S, Khodayari S, Khodayari H, Shahbazi S, Zahedi A, Sohanaki H, Khaniki M. Effects of exercise training together with tamoxifen in reducing mammary tumor burden in mice: Possible underlying pathway of miR-21. European journal of pharmacology. 2015 Oct 15;765:179-87. https://doi.org/10.1016/j.ejphar.2015.08.031.
  12. Bucak MN, Ateşşahin A, Varışlı Ö, Yüce A, Tekin N, Akçay A. The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen: microscopic and oxidative stress parameters after freeze– thawing process. Theriogenology. 2007 Mar 15;67(5):1060-7. https://doi.org/10.1016/j.theriogenology.2006.12.004.
  13. Hatef B, Taromchi A, Nejatbakhsh R, Farrokhi A, Shokri S. Supplementation of freezing media with stromal cell-derived factor1α preserves human sperm from cryodamage. Cryobiology. 2017 Dec 1;79:37-42. https://doi.org/10.1016/j.cryobiol.2017.09.004.
  14. Keogh LM, Byrne PG, Silla AJ. The effect of antioxidants on sperm motility activation in the Booroolong frog. Animal reproduction 2017 Aug 1;183:126-31. https://doi.org/10.1016/j.anireprosci.2017.05.008.
  15. Yan W, Kanno C, Oshima E, Kuzuma Y, Kim SW, Bai H, Takahashi M, Yanagawa Y, Nagano M, Wakamatsu JI, Kawahara M. Enhancement of sperm motility and viability by turmeric by-product dietary supplementation in roosters. Animal reproduction science. 2017 Oct 1;185:195-204. https://doi.org/10.1016/j.anireprosci.2017.08.021.
  16. Mani A, Staikou C, Karmaniolou I, Orfanos N, Mylonas A, Nomikos T, Pafiti A, Papalois A, Arkadopoulos N, Smyrniotis V, Theodoraki N-acetylcysteine and desferoxamine reduce pulmonary oxidative stress caused by hemorrhagic shock in a porcine model. Journal of Investigative Surgery. 2017 Jan 2;30(1):33-40. https://doi.org/10.1080/08941939.2016.1215580.
  17. Santus P, Corsico A, Solidoro P, Braido F, Di Marco F, Scichilone N. Oxidative stress and respiratory system: pharmacological and clinical reappraisal of N-acetylcysteine. COPD: Journal of Chronic Obstructive Pulmonary Disease. 2014 Dec 1;11(6):705-17. https://doi.org/10.3109/15412555.2014.898040.
  18. Mata‐Campuzano M, Álvarez‐Rodríguez M, Alvarez M, Anel L, de Paz P, Garde JJ, Martínez‐Pastor F. Effect of several antioxidants on thawed ram spermatozoa submitted to 37 C up to four hours. Reproduction in domestic animals. 2012 Dec;47(6):907-14. https://doi.org/10.1111/j.1439-0531.2012.01990.x.
  19. Cooper TG, Noonan E, Von Eckardstein S, Auger J, Baker HW, Behre HM, Haugen TB, Kruger T, Wang C, Mbizvo MT, Vogelsong World Health Organization reference values for human semen characteristics. Human reproduction update. 2010 Jan 1;16(3):231- 45. https://doi.org/10.1093/humupd/dmp048.
  20. Kvist U, Björndahl L, editors. Manual on basic semen analysis: 2002. Oxford University Press; 2002. https://doi.org/10.1016/j.fertnstert.2010.01.058.
  21. Garcez ME, dos Santos Branco C, Lara LV, Pasqualotto FF, Salvador Effects of resveratrol supplementation on cryopreservation medium of human semen. Fertility and sterility. 2010 Nov 1;94(6):2118-21. https://doi.org/10.1016/j.fertnstert.2010.01.058.
  22. Buege JA, Aust SD. [30] Microsomal lipid peroxidation. InMethods in enzymology 1978 Jan 1 (Vol. 52, pp. 302-310). Academic Press.
  23. Sá R, Cunha M, Rocha E, Barros A, Sousa M. Sperm DNA fragmentation is related to sperm morphological staining patterns. Reproductive biomedicine online. 2015 Oct 1;31(4):506-15. https://doi.org/10.1016/j.rbmo.2015.06.019.
  24. Rowe PJ, Comhaire FH, Hargreave TB, MahmoudAMA, WHO manual for the standardized investigation and diagnosis of the infertile male. 2000: Cambridge University Press.
  25. Nicholson C, Abramsson L, Holm SE, Bjurulf E, Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using silane-coated silica particles at different g forces. Hum Reprod 2000; 15(3): 662-6.
  26. Fraser L, Strzeżek J. Effects of freezing–thawing on DNA integrity of boar spermatozoa assessed by the neutral comet assay. Reproduction in domestic animals. 2005 Dec;40(6):530-6. https://doi.org/10.1111/j.1439-0531.2005.00626.x.
  27. Sanocka D, Miesel R, Jedrzejczak P, Chełmonska-Soyta AC, Kurpisz M. Effect of reactive oxygen species and the activity of antioxidant systems on human semen; association with male infertility. Int J Androl 1997; 20(5): 255- 64.
  28. O'Connell M, McClure N, Lewis S. The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod 2002; 17(3): 704-9.
  29. Ball BA, Vo A, Detection of lipid peroxidation in equine spermatozoa based upon the lipophilicfluorescent dye C1l-BODIPY581/591. J Androl 2002; 23(2): 259-69.
  30. Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MC. The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. J Androl 2000; 21(6): 895-902.
  31. Lass A, Akagbosu F, and Brinsden P, Sperm banking and assisted reproduction treatment for couples following cancer treatment of the male partner. Hum Reprod Update 2001; 7(4): 370-7.
  32. Donnelly ET, McClure N, Lewis SE. Cryopreservation of human semen and prepared sperm: effects on motility parameters and DNA integrity. Fertil Steril 2001; 76(5): 892-900.
  33. Branco CS, Garcez ME, Pasqualotto FF, Erdtman B, Salvador M. Resveratrol and ascorbic acid prevent DNA damage induced by cryopreservation in human semen. Cryobiology 2010; 60(2): 235-7.
  34. Taylor K, Roberts P, Sanders K, Burton P. Effect of antioxidant supplementation of cryopreservation medium on post-thaw integrity of human spermatozoa. Reprod Biomed Online 2009; 18(2): 184-9.
  35. Garcez ME, dos Santos Branco C, Lara LV, Pasqualotto FF, Salvador M. Effects of resveratrol supplementation on cryopreservation medium of human semen. Fertil Steril 2010; 94(6): 2118-21.
  36. Agarwal A, Prabakaran SA, Said TM. Prevention of oxidative stress injury to sperm. Journal of andrology. 2005 Nov 12;26(6):654-60. https://doi.org/10.2164/jandrol.05016.
  37. Agarwal A, Ikemoto I, Loughlin K. Levels of reactive oxygen species before and after sperm preparation: comparison of swim-up and L4 filtration. Arch Androl 1994; 32: 169.
  38. Chan MM, Mattiacci JA, Hwang HS, Shah A, Fong D. Synergy between ethanol and grape polyphenols, quercetin, and resveratrol, in the inhibition of the inducible nitric oxide synthase pathway. Biochem Pharmacol 2000; 60(10): 1539-48
  39. Shin S, Jeon JH, Park D, Jang MJ, Choi JH, Choi BH, et al. trans-Resveratrol relaxes the corpus cavernosum ex vivo and enhances testosterone levels and sperm quality in vivo. Arch Pharm Res 2008; 31(1): 83-7.
  40. Juan ME, González-Pons E, Munuera T, Ballester J, Rodríguez-Gil JE, Planas JM. transResveratrol, a natural antioxidant from grapes, increases sperm output in healthy rats. J Nutr 2005; 135(4): 757-60.
  41. Lima S, Cedenho A, Cenedeze M, Bertolla R, Oehninger S, Srougi M. Male infertility and heat shock protein 70–2 (HSP70–2) expression. Fertility and Sterility. 2004 Sep 1;82:S181. https://doi.org/10.1016/j.fertnstert.2004.07.475.
  42. Ji ZL, Duan YG, Mou LS, Allam JP, Haidl G, Cai ZM. Association of heat shock proteins, heat shock factors and male infertility. Asian Pacific Journal of Reproduction. 2012 Mar 1;1(1):76-84. https://doi.org/10.1016/S2305-0500(13)60053-6.
  43. Mayer MP, Bukau B. Hsp70 chaperones: cellular functions and molecular mechanism. Cellular and molecular life sciences. 2005 Mar 1;62(6):670. https://doi.org/10.1007/s00018-004-4464-6.
  44. Zhang XG, Hong JY, Yan GJ, Wang YF, Li QW, Hu JH. Association of heat shock protein 70 with motility of frozen-thawed sperm in bulls. Czech Journal of Animal Science. 2015 Jan 1;60(6):256-62. https://doi.org/10.17221/8239-CJAS.
  45. Amidi F, Pazhohan A, Nashtaei MS, Khodarahmian M, Nekoonam S. The role of antioxidants in sperm freezing: a review. Cell and tissue banking. 2016 Dec 1;17(4):745-56. https://doi.org/10.1007/s10561- 016-9566-5.