8954856055505db

توانایی آنتی اکسیدان آنزیمی کاتالاز در بهبود انجماد‌پذیری اسپرم اسب ترکمن

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه علوم دامی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج

2 استاد، گروه علوم دامی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران

3 استادیار، گروه علوم دامی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج و استادیار مؤسسۀ تحقیقات واکسن و سرم سازی رازی، کرج

4 استادیار، مؤسسۀ تحقیقات واکسن و سرم سازی رازی

5 کارشناس علوم آزمایشگاهی، مؤسسۀ تحقیقات واکسن و سرم سازی رازی

6 کارشناس دامپزشکی، مؤسسۀ تحقیقات واکسن و سرم سازی رازی

چکیده

هدف از انجام این آزمایش بررسی تأثیر پاداکسندة (آنتی­اکسیدان) آنزیمی کاتالاز بر بهبود انجمادپذیری اسپرم اسب­های ترکمن بود. در این پژوهش چهار سر نریان بالغ ترکمن با میانگین سنی هشت تا ده سال برای اسپرم­گیری به کار گرفته شد. پس از گرفتن منی و فرآوری آن در آزمایشگاه، اسپرم­های رقیق‌شده به گروه­های تیماری حاوی 0 (شاهد)، 100 (CAT100)، 150 (CAT150) و 200 (CAT200) واحد پاداکسندة کاتالاز در هر میلی­لیتر رقیق‌کنندۀ تقسیم‌شدۀ و با یک روش استاندارد، منجمد شدند. پس از یخ­گشایی برای همۀ تیمارها جنبایی، زنده‌مانی، یکپارچگی غشا، ناهنجاری کل و میزان پراکسیداسیون لیپیدی ارزیابی شد. نتایج نشان داد که افزودن 200 واحد بر میلی­لیتر کاتالاز به رقیق­کنندۀ اسپرم اسب، سبب بهبود فراسنجه‌های جنبایی کل، زنده­مانی، یکپارچگی غشایی و کاهش پراکسیداسیون لیپید، نسبت به گروه شاهد و دیگر گروه­های تیماری شد (05/0P<). همچنین تیمار 150 واحد بر میلی­لیتر کاتالاز نیز سبب بهبود جنبایی کل، زنده­مانی و کاهش پراکسیداسیون لیپید نسبت به گروه شاهد شد (05/0P<). همچنین نتایج نشان می­دهد که افزودن 100 واحد بر میلی­لیتر کاتالاز تأثیر مثبتی روی اغلب فراسنجه‌های کیفی اسپرم ندارد (05/0P>). بر پایۀ نتایج به‌دست‌آمده، تیمارهای آزمایشی تأثیری در کاهش اسپرم‌های نابهنجار نسبت به گروه کنترل نداشتند (05/0P>). درنتیجه می‌توان گفت که افزودن 200 واحد بر میلی‌لیتر کاتالاز به رقیق‌کننده، سبب بهبود انجمادپذیری اسپرم اسب­های ترکمن می­شود.

کلیدواژه‌ها


عنوان مقاله [English]

The potential of catalase as an enzymatic antioxidant to improve freezability of Turkmen stallions sperm

نویسندگان [English]

  • Afshin Seifi Jamadi 1
  • Ahmad Zareh 2
  • Hamid Kohram 3
  • Abolfazl Akbari 4
  • Mohsen Zamen 5
  • Abolfazl Vakhideh 6
1 Ph.D. Student, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
2 Professor, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran
3 Assistant Professor, University College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran and Assistant Professor of Razi Vaccine and Serum Research Institute, Karaj, Iran
4 Assistant Professor, Razi Vaccine and Serum Research Institute, Karaj, Iran
5 Medical Laboratory Expert, Razi Vaccine and Serum Research Institute, Karaj, Iran
6 Veterinary Expert, Razi Vaccine and Serum Research Institute, Karaj, Iran
چکیده [English]

Four mature stallions were used to study the effect of enzymatic antioxidant catalase on freezability of Turkmen stallion's sperm. Collected semen from stallions were processed and pooled before being divided treatments (200×106  sperm/ml) and diluted with extenders supplemented with different levels of catalase [0 (control), 100 U/ml (CAT100), 150U/ml (CAT150) and 200U/ml (CAT200)]. Extended and supplemented semen's were frozen according to a standard protocol. After thawing, motility, viability, membrane integrity, total abnormality and lipid peroxidation were assessed. Results showed that addition of 200U/ml catalase could improve progressive and total motility, viability, membrane integrity and decreased lipid peroxidation compared to control and other levels of catalase (p<0.05). In addition, 150 U/ml catalase improved total motility, viability and decreased MDA concentration (p<0.05). However, the addition of 100 U/ml catalase had no significant effect on sperm quality (p>0.05). The results showed that different levels of catalase could not decreased total abnormality (p>0.05). We concluded that addition of 200U/ml catalase could improve freezability of Turkmen stallion’s sperm.

کلیدواژه‌ها [English]

  • Catalase
  • Free radicals
  • horse semen freezing
  • Lipid Peroxidation
  • Turkmen horse
  1. Agarwal, A. & Sekhon, L.H. (2010). The role of antioxidant therapy in the treatment of male infertility. Human Fertility, 13, 217-225.
  2. Agarwal, A., Sharma, R.K., Sharma, R., Assidi, M., Abuzenadah, A.M., Alshahrani, S., Durairajanayagam, D. & Sabanegh, E. (2014). Characterizing semen parameters and their association with reactive oxygen species in infertile men. Reproductive Biology and Endocrinology 12, 33.
  3. Aitken, R.J., Jones, K.T. & Robertson, S.A. (2012). Reactive oxygen species and sperm function In sickness and in health. Journal of Andrology 33, 1096-1106.
  4. Alvarenga, M.A., Papa, F.O., Carmo, M.T., Kievitsbosch, T., Castro Chaves, M.M.B. & Ramires Neto, C. (2012). Methods of Concentrating Stallion Semen. Journal of Equine Veterinary Science, 32, 424-429.
  5. Asadpour, R., Jafari, R. & Tayefi-Nasrabadi, H. (2012). Effect of various levels of catalase antioxidant in semen extenders on lipid peroxidation and semen quality after the freeze-thawing bull semen. Veterinary Research Forum, pp. 218-221.
  6. Ball, B.A. (2008). Oxidative stress, osmotic stress and apoptosis: Impacts on sperm function and preservation in the horse. Animal Reproduction Science, 107, 257-267.
  7. Baumber, J., Ball, B.A. & Linfor, J.J. (2005). Assessment of the cryopreservation of equine spermatozoa in the presence of enzyme scavengers and antioxidants. American Journal of Veterinary Research, 66, 772-779.
  8. Bouayed, J. & Bohn, T. (2010). Exogenous antioxidants double-edged swords in cellular redox state: health beneficial effects at physiologic doses versus deleterious effects at high doses. Oxidative Medicine and Cellular Longevity, 3, 228-237.
  9. Copeland-Fitzpatrick, J. & Tebay, J. (1998). Hippotherapy and therapeutic riding: An international review. Companion Animals in Human Health, 41-58.
  10. Cristanelli, M., Amann, R., Squires, E. & Pickett, B. (1985). Effects of egg yolk and glycerol levels in lactose-EDTA-egg yolk extender and of freezing rate on the motility of frozen-thawed stallion spermatozoa. Theriogenology, 24, 681-686.
  11. Da Silva, F., Marques, A. & Chaveiro, A. (2010). Reactive oxygen species: a double-edged sword in reproduction. Open Veterinary Science Journal, 4, 127-133.
  12. Evans, G. & Maxwell, W.C. (1987). Salamons' artificial insemination of sheep and goats. (No. Ed. 2). Butterworths.
  13. Fayrer-Hosken, R., Abreu-Barbosa, C., Heusner, G. & Jones, L. (2008). Cryopreservation of stallion spermatozoa with INRA96 and glycerol. Journal of Equine Veterinary Science, 28, 672-676.
  14. Fernández‐Santos, M.R., Martínez‐Pastor, F., García‐Macías, V., Esteso, M.C., Soler, A.J., Paz, P., Anel, L. & Garde, J.J. (2007). Sperm characteristics and DNA integrity of Iberian red deer (Cervus elaphus hispanicus) epididymal spermatozoa frozen in the presence of enzymatic and nonenzymatic antioxidants. Journal of Andrology, 28, 294-305.
  15. Gibb, Z., Butler, T., Morris, L., Maxwell, W. & Grupen, C. (2013). Quercetin improves the postthaw characteristics of cryopreserved sex-sorted and nonsorted stallion sperm. Theriogenology, 79(6), 1001-1009.
  16. Holland, M.K., Alvarez, J.G. & Storey, B.T. (1982). Production of superoxide and activity of superoxide dismutase in rabbit epididymal spermatozoa. Biology of Reproduction, 27, 1109-1118.
  17. Holt, W. (2000). Basic aspects of frozen storage of semen. Animal Reproduction Science, 62, 3-22.
  18. Kefer, J.C., Agarwal, A. & Sabanegh, E. (2009). Role of antioxidants in the treatment of male infertility. International Journal of Urology, 16, 449-457.
  19. Kirk, E., Squires, E. & Graham, J. (2005). Comparison of in vitro laboratory analyses with the fertility of cryopreserved stallion spermatozoa. Theriogenology, 64, 1422-1439.
  20. Kmenta, I., Strohmayer, C., Müller-Schlösser, F. & Schäfer-Somi, S. (2011). Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of cold-stored canine spermatozoa. Theriogenology, 75, 1095-1103.
  21. Li, Z., Lin, Q., Liu, R., Xiao, W. & Liu, W. (2010). Protective effects of ascorbate and catalase on human spermatozoa during cryopreservation. Journal of Andrology, 31, 437-444.
  22. Loomis, P.R. & Squires, E.L. (2005). Frozen semen management in equine breeding programs. Theriogenology, 64, 480-491.
  23. Love, C., Thompson, J., Brinsko, S., Rigby, S., Blanchard, T., Lowry, V. & Varner, D. (2003). Relationship between stallion sperm motility and viability as detected by two fluorescence staining techniques using flow cytometry. Theriogenology, 60, 1127-1138.
  24. Luo, S.-M., Schatten, H. & Sun, Q. Y. (2013). Sperm mitochondria in reproduction: good or bad and where do they go? Journal of Genetics and Genomics, 40, 549-556.
  25. Michael, A., Alexopoulos, C., Pontiki, E., Hadjipavlou-Litina, D., Saratsis, P. & Boscos, C. (2007). Effect of antioxidant supplementation on semen quality and reactive oxygen species of frozen-thawed canine spermatozoa. Theriogenology, 68, 204-212.
  26. Partyka, A., Łukaszewicz, E. & Niżański, W. (2012). Lipid peroxidation and antioxidant enzymes activity in avian semen. Animal Reproduction Science, 134, 184-190.
  27. Revell, S.G. & Mrode, R.A. (1994). An osmotic resistance test for bovine semen. Animal Reproduction Science, 36(1), 77-86.
  28. Rossi, T., Mazzilli, F., Delfino, M. & Dondero, F. (2001). Improved human sperm recovery using superoxide dismutase and catalase supplementation in semen cryopreservation procedure. Cell and Tissue Banking, 2, 9-13.
  29. Salazar Jr, J., Teague, S., Love, C., Brinsko, S., Blanchard, T. & Varner, D. (2011). Effect of cryopreservation protocol on postthaw characteristics of stallion sperm. Theriogenology, 76, 409-418.
  30. Sanocka, D. & Kurpisz, M. (2004). Reactive oxygen species and sperm cells. Reproductive Biology and Endocrinology, 2, 1-7.
  31. Seifi-Jamadi, A., Kohram, H., Zareh-Shahne, A., Dehghanizadeh, P. & Ahmad, E. (2016). Effect of various concentrations of butylated hydroxyanisole and butylated hydroxytoluene on freezing capacity of Turkman stallion sperm. Animal Reproduction Science, 170, 108-13.
  32. Seifried, H. E., Anderson, D. E., Milner, J. A. & Greenwald, P. (2006). Reactive oxygen species and dietary antioxidants: double-edged swords. New developments in antioxidant research. Hauppauge, NY: Nova Science Publishers Inc, 1-25.
  33. Sikka, S.C., Rajasekaran, M. & Hellstrom, W.J. (1995). Role of oxidative stress and antioxidants in male infertility. Journal of Andrology, 16, 464-468.
  34. Squires, E. (2005). Integration of future biotechnologies into the equine industry. Animal Reproduction Science, 89, 187-198.