Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Effects of Arsenite and Antioxidants on Sperm Motility, Plasma Membrane Integrity, Mitochondrial Activity, and Lipid Peroxidation in Pigs

돼지정자의 운동성, 원형질막 온전성, 미토콘드리아 기능성 및 원형질막 지질과산화에 미치는 arsenite 및 항산화제의 영향

  • Kim, Han-Su (College of Animal Life Sciences, Kangwon National University) ;
  • Lee, Yu-Sub (College of Animal Life Sciences, Kangwon National University) ;
  • Lee, Sang-Hee (Institute of Animal Resources, Kangwon National University) ;
  • Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University) ;
  • Lee, Seunghyung (College of Animal Life Sciences, Kangwon National University) ;
  • Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University)
  • 김한수 (강원대학교 동물생명과학대학) ;
  • 이유섭 (강원대학교 동물생명과학대학) ;
  • 이상희 (강원대학교 동물자원공동연구소) ;
  • 정희태 (강원대학교 수의과대학) ;
  • 박춘근 (강원대학교 동물생명과학대학) ;
  • 이승형 (강원대학교 동물생명과학대학) ;
  • 양부근 (강원대학교 동물생명과학대학)
  • Received : 2016.10.26
  • Accepted : 2017.01.25
  • Published : 2017.05.30
Download PDF
⟨ Previous Next ⟩

Abstract

This study was to investigate the role of antioxidants on the characteristics of arsenite-damaged boar semen. Collected sperm was diluted with semen extender, and $100{\mu}M$ arsenite was used for sperm damage. Then melatonin, silymarin, curcumin, and vitamin E were applied for 3, 6, and 9 hr in arsenite-treated boar sperm. Sperm characteristics were then analyzed for motility, plasma membrane integrity, mitochondrial activity, and lipid peroxidation. In the results, sperm motility (control, $77.3{\pm}1.8%$) was decreased by arsenite ($33.3{\pm}1.5%$), while the antioxidant treatment groups (100 nM melatonin, $55.8{\pm}3.4%$; $2{\mu}M$ silymarin, $48.8{pm}3.4%$; $10{\mu}M$ curcumin, $53.9{\pm}2.8%$; and $500{\mu}M$ vitamin E, $54.5{\pm}3.1%$) showed increases compared to the arsenite group (p<0.05). $100{\mu}M$ arsenite decreased the sperm plasma membrane integrity ($24.5{\pm}1.6%$) and mitochondrial activity ($58.2{\pm}2.6%$), and increased lipid peroxidation ($5.3{\pm}0.2%$) at 3 hr (p<0.05). However, arsenite-treated samples with 100 nM melatonin, $2{\mu}M$ silymarin, $10{\mu}M$ curcumin, and $500{\mu}M$ vitamin E increased the plasma membrane integrity and mitochondria activity, and decreased lipid peroxidation compared to the arsenite-treated samples. In summary, arsenite may induce sperm damage and oxidation stress, while antioxidants such as melatonin, silymarin, curcumin, and vitamin E are useful for maintaining sperm characteristics. Therefore, antioxidants can protect sperm against damage by arsenite in fresh boar semen.

본 실험은 항산화제인 melatonin, silymarin, curcumin 및 vitamin E가 aresenite에 의해 손상된 돼지의 정자성상능력에 미치는 영향을 조사한 연구이다. 돼지정자는 채취 후 희석하여 실험에 이용하였으며, $100{\mu}M$ arsenite는 인위적으로 정자를 손상시키는데 사용하였다. 또한 100 nM melatonin, $2{\mu}M$ silymarin, $10{\mu}M$ curcumin 및 $500{\mu}M$ Vitamin E를 희석된 돼지정자에 첨가하여 3, 6 및 9시간 동안 배양 후 정자의 운동성, 원형질막 온전성, 미토콘드리아 기능성 및 원형질막 지질과산화를 검토하였다. 그 결과, 정자의 운동성 및 원형질막 온전성은 $100{\mu}M$ arsenite 처리구에서 유의적으로 감소하였으며, 항산화제 단독처리구는 유의적으로 증가하였다. 또한, arsenite에 의해 손상된 처리구는 항산화제에 의해 유의적으로 증가하였다(p<0.05). 정자의 미토콘드리아 기능성은 $100{\mu}M$ arsenite 처리구에서 유의적으로 감소하였고, 정자의 원형질막 지질과산화는 $100{\mu}M$ arsenite 처리구에서 유의적으로 증가하는 것으로 나타났다(p<0.05). 결론적으로, arsenite의해 감소된 돼지정자의 운동성과 원형질막은 melatonin, silymarin, curcumin 및 vitamin E와 같은 항산화제에 의하여 예방될 수 있다고 판단되며, arsenite에 의해 감소된 정자의 수정능력은 항산화제에 의해 회복될 수 있을 것이라 생각된다.

Keywords

References

  1. Aitken, R. J. and Roman, S. D. 2008. Antioxidant systems and oxidative stress in the testes. Oxi. Med. Cell Longev. 1, 15-24. https://doi.org/10.4161/oxim.1.1.6843
  2. Ashrafi, I., Kohram, H. and Ardabili, F. F. 2013. Antioxidative effects of melatonin on kinetics, microscopic and oxidative parameters of cryopreserved bull spermatozoa. Anim. Reprod. Sci. 139, 25-30. https://doi.org/10.1016/j.anireprosci.2013.03.016
  3. Bashandy, S. A., ElAwdan, S. A., Ebaid, H. and Alhazza, I. M. 2016. Antioxidant potential of spirulina platensis mitigates oxidative stress and reprotoxicity induced by sodium arsenite in male rats. Oxi. Med. Cell Longev. 2016, 1-8.
  4. Basu, A., Mahata, J., Gupta, S. and Giri, A. k. 2001. Genetic toxiCOlogy of a paradoxical human carcinogen, arsenic: a review. Mutat. Res. 488, 171-194. https://doi.org/10.1016/S1383-5742(01)00056-4
  5. Celino, F. T., Yamaguchi, S., Miura, C. and Miura, T. 2009. Arsenic inhibits in vitro spermatogenesis and induces germ cell, apoptosis in Japanese eel (Anguilla japonica). Reproduction 138, 279-287. https://doi.org/10.1530/REP-09-0167
  6. Chitra, K. C., Latchoumycandane, C. and Mathur, P. P. 2003. Induction of oxidative stress by bisphenol A in the epididymal sperm of rats. Toxicology 185, 119-127. https://doi.org/10.1016/S0300-483X(02)00597-8
  7. Das, S., Upadhaya, P. and Giri, S. 2016. Arsenic and smokeless tobacCO induce genotoxicity, sperm abnormality as well as oxidative stress in mice in vivo. Genes Environ. 38, 1-4. https://doi.org/10.1186/s41021-016-0028-x
  8. Eskandari, F. and Momeni, H. R. 2016. Protective effect of silymarin on viability, Motility and mitochondrial membrane potential of ram sperm treated with sodium arsenite. Int. J. Reprod. Biomed. 14, 397-402. https://doi.org/10.29252/ijrm.14.6.397
  9. Ferreira, M., Matos, R. C., Oliveira, H., Nunes, B. and de Lourdes Pereira, M. 2012. Impairment of mice spermatogenesis by sodium arsenite. Hum. Exp. Toxicol. 31, 290-302. https://doi.org/10.1177/0960327111405862
  10. Flora, S. J. 2011. Arsenic-induced oxidative stress and its reversibility. Free. Radic. Biol. Med. 51, 257-281. https://doi.org/10.1016/j.freeradbiomed.2011.04.008
  11. Flora, S.J. 2016. Arsenic and dichlorvos: Possible interaction between two environmental contaminants. J. Trace Elem. Med. Biol. 35, 43-60. https://doi.org/10.1016/j.jtemb.2016.01.012
  12. Gharagozloo, P. and Aitken, R. J. 2011. The role of sperm oxidative stress in male infertility and the significance of oral antioxidant therapy. Hum. Reprod. 26, 1628-1640 https://doi.org/10.1093/humrep/der132
  13. Guan, X., Du, J., Meng, X., Sun, Y., Sun, B. and Hu, Q. 2012. Application of titanium dioxide in arsenic removal from water : a review. J. Hazard Mater. 215-216, 1-16 https://doi.org/10.1016/j.jhazmat.2012.02.069
  14. Hong, Y. S., Song, K. H. and Chung, J. Y. 2014. Health effects of chronic arsenic exposure. J. Prev. Med. Public Health 47, 245-252. https://doi.org/10.3961/jpmph.14.035
  15. Jang, H. Y., Jin, H. A., Lee, H. Y., Kim, D. J., Cheong, H. T., Kim, J. T., Park, I. C., Park, C. K. and Yang, B. K. 2009. Silymarin and Vit E alleviate alone or synergetically hydrogen peroxide induced-oxidative stress on boar sperm characteristics during in vitro storage. Reprod. Dev. Biol. 33, 273-281.
  16. Jang, H. Y., Kim, Y. H., Cheong, H. T., Kim, J. T., Park, I. C., Park, C. K. and Yang, B. K. 2009. Silymarin attenuates hydrogen peroxide induced oxidative stress on semen characteristics during in vitro storage of boar semen. Reprod. Dev. Biol. 33, 99-105.
  17. Jang, H. Y., Kim, Y. H., Kim, B. W., Park, I. C., Cheong, H. T., Kim, J. T., Park, C. K., Kong, H. S., Lee, H. K. and Yang, B. K. 2010. Ameliorative effects of melatonin against hydrogen peroxide‐induced oxidative stress on boar sperm characteristics and subsequent in vitro embryo development. Reprod. Domest. Anim. 45, 943-950. https://doi.org/10.1111/j.1439-0531.2009.01466.x
  18. Jeon, Y. J. and Kim, D. Y. 2013. The antioxidant effect of silymarin on miniature pig sperm cryopreservation. J. Chitin. Chitosan. 18, 190-197.
  19. Lengfelder, E., Hofmann, W. K. and Nowak, D. 2012. Impact of arsenic trioxide in the treatment of acute promyelocytic leukemia. Leukemia 26, 433-442. https://doi.org/10.1038/leu.2011.245
  20. Liu, Q., Zhou, Y. F., Duan, R. J., Wei, H. K., Peng, J. and Jiang, S. W. 2016. Dietary n-6:n-3 ratio and Vitamin E improve motility characteristics in association with membrane properties of boar spermatozoa. Asian J. Androl. 18, 1-7.
  21. Marchetti, C., Obert, G., Deffosez, A., Formstecher, P. and Marchetti, P. 2002. Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm. Hum. Reprod. 17, 1257-1265. https://doi.org/10.1093/humrep/17.5.1257
  22. Mayorga-Torresa, B. J., Cardona-Mayaa, W., Cadavid, A. and Camargo, M. 2013. Evaluation of sperm functional parameters in normozoospermic infertile individuals. Actas. Urol. Esp. 37, 221-227. https://doi.org/10.1016/j.acuro.2012.06.008
  23. Mazur, P., Katkova, I. I., Katkova, N. and Critser, J. K. 2000. The enhancement of the ability of mouse sperm to survive freezing and thawing by the use of high concentrations of glycerol and the presence of an escherichia coli membrane preparation (oxyrase) to lower the oxygen concentration. Cryobiology 40, 187-209. https://doi.org/10.1006/cryo.2000.2238
  24. Momeni, H. R. and Eskandari, N. 2012. Effect of vitamin E on sperm parameters and DNA integrity in sodium arsenite- treated rats. Iran J. Reprod. Med. 10, 249-256.
  25. Momeni, H. R., Oryan, S. and Eskandari, N. 2012. Effect of vitamin E on sperm number and testis histopathology of sodium arsenite-treated rats. Reprod. Biol. 12, 171-181. https://doi.org/10.1016/S1642-431X(12)60084-9
  26. Muratorl, M., Maggi, M., Spinelli, S., Filimberti, E., Forti, G. and Baldi, E. 2003. Spontaneous DNA fragmentation in swim-up selected human spermatozoa during long term incubation. J. Androl. 24, 253-262. https://doi.org/10.1002/j.1939-4640.2003.tb02670.x
  27. Pant, N., Kumar, R., Murthy, R. C. and Srivastava, S. P. 2001. Male reproductive effect of arsenic in mice. Biometals 14, 113-117. https://doi.org/10.1023/A:1016686113763
  28. Pant, N., Murthy, R. C. and Srivastava, S. P. 2004. Male reproductive toxicity of sodium arsenite in mice. Hum. Exp. Toxicol. 23, 399-403. https://doi.org/10.1191/0960327104ht467oa
  29. Reddya, P. S., Rania, G. P., Sainatha, S. B., Meenaa, R. and Supriyaa, C. H. 2011. Protective effects of N-acetylcysteine against arsenic-induced oxidative stress and reprotoxicity in male mice. J. Trace. Elem. Med. Biol. 25, 247-253. https://doi.org/10.1016/j.jtemb.2011.08.145
  30. Roostaei-Ali Mehr, M. and Parisoush, P. 2016. Effect of different levels of silymarin and caproic acid on storage of ram semen in liquid form. Reprod. Domest. Anim. 51, 569-574. https://doi.org/10.1111/rda.12721
  31. Sarkar, M., Chaudhuri, G. R., Chattopadhyay, A. and Biswas, N. M. 2003. Effect of sodium arsenite on spermatogenesis, plasma gonadotrophins and testosterone in rats. Asian J. Androl. 5, 27-32.
  32. Sengupta, A., Ghosh, M. and Bhattacharyya, D. K. 2014. Antioixidative effect of rice bran oil and medium chain fatty acid rich rice bran oil in arsenite induced oxidative stress in rats. J. Oleo. Sci. 63, 1117-1124. https://doi.org/10.5650/jos.ess14128
  33. Tokar, E. J., Diwan, B. A., Ward, J. M., Delker, D. A. and Waalkes, M. P. 2011. Carcinogenic effects of "Whole-Life" exposure to inorganic arsenic in CD1 mice. Toxicol. Sci. 119, 73-83. https://doi.org/10.1093/toxsci/kfq315
  34. Tvrda, E., Lukac, N., LukaCOva, J., Knazicka, Z. and Massanyi, P. 2013. Stimulating and protective effects of vitamin E on bovine spermatozoa. JMBFS 2, 1386-1395.
  35. Vernet, P., Aitken, R. J. and Drevet, J. R. 2004. Antioxidant strategies in the epididymis. Mol. Cell. Endocrinol. 216, 31-39. https://doi.org/10.1016/j.mce.2003.10.069
  36. Vernet, P., Fulton, N., Wallace, C. and Aitken, R. J. 2001. Analysis of reactive oxygen species generating systems in rat epididymal spermatozoa. Biol. Reprod. 65, 1102-1113. https://doi.org/10.1095/biolreprod65.4.1102
  37. Wang, J. P., Qi, L., Moore, M. R. and Ng, J. C. 2002. A review of animal models for the study of arsenic carcinogenesis. Toxcol. Lett. 133, 17-31. https://doi.org/10.1016/S0378-4274(02)00086-3
  38. Wright, C., Milne, S. and Leeson, H. 2014. Sperm DNA damage caused by oxidative stress: modifiable clinical, lifestyle and nutritional factors in male infertility. Reprod. Biomed. Online 28, 684-703. https://doi.org/10.1016/j.rbmo.2014.02.004

Cited by

  1. Ameliorative Effect of Chitosan Complex on Miniature Pig Sperm Cryopreservation vol.33, pp.4, 2018, https://doi.org/10.12750/JET.2018.33.4.337