DOI QR코드

DOI QR Code

Effects of Endocrine Disruptors (NP, DBP and BPA) on Sperm Characteristics and Development of IVF Embryos in Pig

  • Yuh, In Suh (College of Animal Life Science, Kangwon National University) ;
  • Cheong, Hee Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Kim, Jong Taek (School of Veterinary Medicine, Kangwon National University) ;
  • Park, In Chul (School of Veterinary Medicine, Kangwon National University) ;
  • Park, Choon Keun (College of Animal Life Science, Kangwon National University) ;
  • Yang, Boo Keun (College of Animal Life Science, Kangwon National University)
  • Received : 2013.06.27
  • Accepted : 2013.08.07
  • Published : 2013.08.31

Abstract

This study was to examine single or combined in vitro effects of environmental endocrine disruptors on boar sperm characteristics, oxidative stress damage in sperm and development of porcine IVF embryos. Addition of various concentration of NP (10, 20, $30{\mu}M$), DBP (10, 50, $100{\mu}M$) and BPA (1, 5 or $10{\mu}g/ml$) on boar sperm characteristics such as percentages of sperm motility, viability, membrane integrity and mitochondrial activity were dose-dependently decreased within 3, 6 or 9 hr incubation period (p<0.05). The overall detrimental effects increased with incubation time increasement. NP, DBP and BPA showed the detrimental effects on sperm membrane and mitochondria of energy production organelles affecting cell viability with the dependancy of dose and incubation time. In combination effects, NP ($10{\mu}M$) + DBP ($10{\mu}M$) significantly decreased boar general sperm characteristics for 3 or 6 hr incubation period compared with control (p<0.05). When both of NP and DBP concentrations (NP; $30{\mu}M$, DBP; $100{\mu}M$) increase, the detrimental effects on sperm characteristics were larger than those of low concentration combination (p<0.05). The inhibitory effects of NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$) on sperm characteristics were larger than those of NP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) (p<0.05). DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) decreased sperm characteristics compared with the low concentration combination (DBP $10{\mu}M$ + BPA $1{\mu}g/ml$, p<0.05). This result indicates the detrimental effects of both chemicals on sperm characteristics were dose dependent. Addition of NP ($30{\mu}M$) + DBP ($100{\mu}M$), NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$), DBP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) or DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) significantly increased lipid peroxidation for 3 or 6 hr incubation period (p<0.05) compared with no addition control. NP (${\geq}20{\mu}M$) decreased the percentages of IVF embryo development from morulae and blastocyst stages (p<0.05) and its detrimental effects were dose-dependant. BPA 0, 1, 5 or $10{\mu}g/ml$ decreased significantly and dose-dependently the percentage of morulae plus and blastocysts (p<0.05). Combinations of DBP ($100{\mu}M$) plus NP ($30{\mu}M$) and DBP ($100{\mu}M$) plus BPA ($10{\mu}g/ml$) did not affect on morulae and blastocyst development, but NP ($30{\mu}M$) plus BPA ($10{\mu}g/ml$) has significant detrimental effect on embryo development at these stages (p<0.05). These overall results indicate that the partial detrimental effects on boar sperm characteristics and embryo development by NP, DBP, BPA or the combination of these chemicals might be due to the increasement of lipid peroxidation and free radical formation in the cell and there were no specific interaction effects on boar sperm and embryo degeneration among the combined treatments.

Keywords

NP;DBP;BPA;Endocrine disruptors;Sperm characteristics;IVF embryo development

Acknowledgement

Supported by : Kangwon National University

References

  1. Akingbemi, BT, Klinfelter, GR, Zirkin, BR. and Hardy, MP. 2004. Phthalate induced leydig cell hyperplasia associated with multiple endocrine disturbances. Proc Natl Acad Sci USA. 101, 775-780. https://doi.org/10.1073/pnas.0305977101
  2. Bao, AM, Man, XM, Guo, XJ, Dong, HB, Wang, FQ, Sun, H, Wang, YB, Zhou, ZM. and Sha, JH. 2011. Effects of di-n-butyl phthalate on male rat reproduction following pubertal exposure. Asian J Andro. 13, 702-709. https://doi.org/10.1038/aja.2011.76
  3. Boockfor, FR. and Blake, CA. 1997. Chronic administration of 4-tert-octylphenol to adult male rats causes shrinkage of the testes and male accessory sex organs, disrupts spermatogenesis, and increase the incidence of sperm deformities. Biol Reprod. 57, 267-277. https://doi.org/10.1095/biolreprod57.2.267
  4. Bolt, HM, Janninq, P, Michna, H. and Degen, GH. 2001. Comparative assessment of endocrine modulators with oestrogenic activity: I. Definition of a hygiene-based margein of safety (HBMOS) for xeno-oestrogens against the background of European developments. Arch Toxicol. 74, 649-662. https://doi.org/10.1007/s002040000178
  5. Braun, JM, Smith, KW, Williams, PL, Calafat, AM, Berry, K, Ehrlich, S. and Hauser, R. 2012. Variability of urinary phthalate metabolite and bisphenol A concentrations before and during pregnancy. Environ Health Perspect. 120, 739-745. https://doi.org/10.1289/ehp.1104139
  6. Brian, JV, Harris, CA, Scholze, M., Backhaus, T, Booy, P., Lamoree. 2005. Accurate prediction of the response of freshwater fish to a mixture of estrogenic chemicals. Environ Health Prospect. 113, 721-728. https://doi.org/10.1289/ehp.7598
  7. El-Dakdoky, MH. and Helal, MA. 2007. Reproductive toxicity of male mice after exposure to nonylphenol. Bull Environ Contam Toxicol. 79, 188-191. https://doi.org/10.1007/s00128-007-9158-y
  8. Fabjan, E, Hulzebos, E, Mennes, W. and Piersm, AH. 2006. A category approach for reproductive effects of phthalates. Crit Rev Toxicol. 36, 695-726. https://doi.org/10.1080/10408440600894914
  9. Fisher, JS, Macpherson, S. and Marchetti, N. 2003. Human 'testicular dysgenesis syndrome'; a possible model using in-utero exposure of the rat to dibutyl phthalate. Hum Reprod. 18, 1383-1394. https://doi.org/10.1093/humrep/deg273
  10. Gang, Y. and Han, XD. 2006. Nonylphenol-induced oxidative stress and cytotoxicity in testicular Sertoli cells. Reprod Toxicol. 22;623-630. https://doi.org/10.1016/j.reprotox.2006.04.019
  11. Gong, Y., Wu, J., Huang, Y., Shen, S. and Han, X. 2009. Nonylphenol induces apoptosis in rat testicular Sertoli cells via endoplasmic reticulum stress. Toxicol Lett. 186, 84-95. https://doi.org/10.1016/j.toxlet.2009.01.010
  12. Gray, LE Jr, Wilson, VS, Stoker, T., Lambright, C. and Furr, J. 2006. Adverse effects of environmental antiandrogens and androgens on reproductive development in mammals. Int J Androl. 29, 96-104. https://doi.org/10.1111/j.1365-2605.2005.00636.x
  13. Guenther, K., Heinke, V., Thiele, B., Kleist, E., Prast, H. and Raecker, T. 2002. Endocrine disrupting nonylphenol are ubiquitous in food. Environ Sci Technol. 36, 1676-1680. https://doi.org/10.1021/es010199v
  14. Hallmark, N., Walker, M., Mckinnell, C., Mahood, IK., Scott, H. and Bayne, R. 2007. Effects of monobuthyl and di (n-butyl) phthalate in vitro on steroidgenesis and leydig cell aggregation in fatal explants from the rats: comparison with effects in vivo in the fatal rat and neonatal marmoset and in vitro in the human. Environ Health Prospect. 115, 390-396.
  15. Han, XD, Tu, ZG, Gong, Y., Shen, SN., Wang, XY. and Kang, LN. 2004. The toxic effects of nonylphenol on the reproductive system of male rats. Reprod Toxicol. 19, 215-221. https://doi.org/10.1016/j.reprotox.2004.06.014
  16. Herath, CB., Jin, W., Watanabe, G., Arai, K., Sukuki, AK. and Taya, K. 2004. Adverse effects of environmental toxicants,octylphenol and bisphenol A, on male reproductive functions in pubertal rats. Endocrine. 25, 163-172. https://doi.org/10.1385/ENDO:25:2:163
  17. Howdeshell, KL., Hotchkiss, AK., Thayer, KA., Vandenbergh, JG. and Vom Saal, FS. 1999. Environmental toxins-exposure to bisphenol A advances puberty. Nature. 401, 763-764. https://doi.org/10.1038/44517
  18. Hunt, PA., Koehler, KE., Susiarjo, M., Hodges, CA., Ilagan, A. and Voigt, RC. 2003. Bisphenol A exposure causes meistic aneuploidy in the female mouse. Curr Biol. 13, 546-553. https://doi.org/10.1016/S0960-9822(03)00189-1
  19. Ichimura, T., Kawamure M. and Mitani A. 2003. Co-localized expression of Fas L, Fas capase 3 and apoptosis DNA fragmentation in mouse testis after oral exposure to di-(2-ethyl hexyl) phthalate. Toxicology. 194, 35-42. https://doi.org/10.1016/j.tox.2003.07.003
  20. Kwack, SJ., Kwon, O., Kim, HS., Kim, SS., Kim, SH. and Sohn, KH. 2002. Comparative evaluation of alkylphenolic compounds on estrogenic activity in vivo and in vitro. J. Toxicol. Environ. Health Part A. 65, 419-431. https://doi.org/10.1080/15287390252808082
  21. Lee, PC., Arndt, P. and Nickels, KC. 1999. Testicular abnormalities in male rats after lactational exposure to nonylphenols. Endocrine. 11, 61-68. https://doi.org/10.1385/ENDO:11:1:61
  22. Li, D., Hu, Y., Shen, X., Dai xinjue, and Han, X. 2010. Combined effects of two environmental endocrine disruptors nonylphenol and di-n-butyl phthalate on rat sertoli cells in vitro. Reprod Toxicol. 30, 438-445 https://doi.org/10.1016/j.reprotox.2010.06.003
  23. Lu, YY., Chen, ML., Sung, FC., Wang, PSG. and Mao, IF. 2007. Daily intake of 4-nonylphenol. Environ Int. 33, 903-910. https://doi.org/10.1016/j.envint.2007.04.008
  24. Mylchreest, E., Sar, M., Cattley, RC. and Foster, OM. 1999. Disruption of androgen-regulated male reproductive development by di (n-butyl) phthalate during late gestation in rats is different from flutamide. Toxicol Appl Pharmacol. 156, 81-95. https://doi.org/10.1006/taap.1999.8643
  25. Pan, G., Hanaoka, T., Yoshimura, M., Zhang, S. and Wang, P. 2006. Decreased serum free testosterone in workers exposed to high levels of di-butyl phthalate (DBP) and Di-2-ethyl hexyl phthalate (DEHP); a cross-sectional study in china. Environ Health Prospect. 114, 1643-1648.
  26. Petro, EML., Leroy, JLMR., Van Cruchten, SJM., Covaci, A., Jorssen, EPA, Bols, PEJ. 2012. Endocrine disruptors and female fertility: Focus on (bovine) ovarian follicular physiology. Theriogenology. 78, 1887-1900. https://doi.org/10.1016/j.theriogenology.2012.06.011
  27. Rastogi, SK. 2006. Phthalate exposure and health outcomes. Indian J Occup Environ Med. 10, 111-115. https://doi.org/10.4103/0019-5278.29570
  28. Tinwell, H. and Ashby, J. 2004. Sensitivity of the immature rat uterotrophic assay to mixture estrogens. Environ. Health Prospect. 112, 575-582. https://doi.org/10.1289/ehp.6831
  29. Uguz, C, Varisli O, Agca, C. and Agca, Y. 2009. Effects of nonylphenol on motility and subcellular elements of epididymal rat sperm. Reprod Toxicol. 28, 542-549. https://doi.org/10.1016/j.reprotox.2009.06.007
  30. Xu, J., Li, J., Feng, Z., Gong, L., Zhang, B. and Yu, J. 2013. Neurotoxic effects of nonylphenol: a review. The central Eur J Med. 125, 61-70.
  31. Yang, DK. and Ding, WH. 2005. Determination of alkylphenolic residues in fresh fruits and vegetables by extractive steam distillation and gas-chromatography-mass spectrometry. J Chromatogr. 1088, 200-204. https://doi.org/10.1016/j.chroma.2004.11.063
  32. Ying, GG., Willians, B. and Kookana R. 2002. Environmental fate of alkylphenols and alkylphenol ethoxylates. A review Environ Int. 28, 215-226. https://doi.org/10.1016/S0160-4120(02)00017-X
  33. Zafra-Gomez, A., Ballesteros, O., Navalon, A. and Vilchez, JL. 2008. Determination of some endocrine disruptor chemicals in urban wastewater samples using liquid chromatography-mass spectrometry. Microchem J. 88, 87-94. https://doi.org/10.1016/j.microc.2007.10.003
  34. Zhang, Y., Jiang, X. and Chen, B. 2004. Reproductive and developmental toxicity in F1 Sprague-Dawlay male rats exposed to di-n-butyl phthalate in utero and during laction and determination of its NOAEL. Reprod Toxicol. 18, 669-676. https://doi.org/10.1016/j.reprotox.2004.04.009