Journal of Life Science (생명과학회지)

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

DOI QR Code

Establishment of Quantitative Evaluation Method for Screening Testicular Toxicity in Rats: 2-Bromopropane as an Example

랫드에서 고환독성의 정색을 위한 정량적 평가법의 확립: 2-bromopropane의 예

  • Cha Shin-Woo (Korea Institute of Toxicology, KRICT) ;
  • Bae Joo-Hyun (Korea Institute of Toxicology, KRICT) ;
  • Son Woo-Chan (Huntingdon Life Sciences) ;
  • Shin Jin-Young (College of Veterinary Medicine, Chonnam National University) ;
  • Shin Dong-Ho (College of Veterinary Medicine, Chonnam National University) ;
  • Kim Sung-Ho (College of Veterinary Medicine, Chonnam National University) ;
  • Park Seung-Chun (College of Veterinary Medicine, Kyungpook National University) ;
  • Kim Jong-Choon (College of Veterinary Medicine, Chonnam National University)
  • Published : 2005.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

The aims of the study were to establish a short-term screening test for detecting testicular toxicity of chemicals in rats and to determine whether a 2-week administration period is sufficient to detect testicular toxicity of 2-bromopropane (2-BP) as an example. Male Sprague-Dawley rats were subcutaneously administered with 1000 mg/kg/day of 2-BP or its vehicle for 2 weeks. Ten male rats each were sacrificed on days 3, 7 and 14 after the initiation of treatment. Parameters of testicular toxicity included genital organ weights, testicular sperm head counts, epididymal sperm counts, motility and morphology, and qualitative and quantitative histopathologic examinations. The early histopathological changes observed on day 3 of treatment included degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, and decreased number of spermatogonia in stages II and V. On day 7 of treatment, atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in stages VII and XII. On day 14 after treatment, a significant decrease in the weights of testes and seminal vesicles was found. Atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in all spermatogenic stages were also observed. In addition, a slight non-significant decrease in testicular sperm head counts, daily sperm production rate and epididymal sperm counts was found. The results showed that 2 weeks of treatment is sufficient to detect the adverse effects of 2-BP on male reproductive organs. It is considered that the short-term testicular toxicity study established in this study can be a useful tool for screening the testicular toxic potential of new drug candidates in rats.

Keywords

References

  1. Bailey, S. A., R. H. Zidell and R. W. Perry. 2004. Relationships between organ weight and body/brain weight in the rat: what is the best analytical endpoint? Toxicol. Pathol. 32, 448-466 https://doi.org/10.1080/01926230490465874
  2. Boekelheide, K., S. P. Darney, G. P. Daston, R. M. David, U. Luderer, A. F. Olshan, W. T. Sanderson, C. C. Willhite and S. Woskie. 2004. NTP-CERHR Expert Panel Report on the reproductive and developmental toxicity of 2-bromopropane. Reprod. Toxicol. 18, 189-217 https://doi.org/10.1016/j.reprotox.2003.10.003
  3. Chung, M. K., S. J. Lee, Y. B. Kim, S. C. Park, D. H. Shin, S. H. Kim and J. C. Kim. 2005. Evaluation of spermatogenesis and fertility in F1 male rats after in utero and neonatal exposure to extremely low frequency electromagnetic fields. Asian J. Androl. 7, 189-194 https://doi.org/10.1111/j.1745-7262.2005.00007.x
  4. Chun, Y. H., J. H. Han and J. J. Yu. 2001. Reproductive toxicity assessment on 2-bromopropane using spermatogenesis stage classification and Sertoli cell indices. J. Toxicol. Pub. Health. 17, 267-272
  5. Chun, M. K. and J. C. Kim. 2000. Principles and methods for the reproductive-toxicological evaluation of new drug candidates. J. Toxicol. Pub. Health 16, 229-238
  6. Creasy, D. M. 1997. Evaluation of testicular toxicity in safety evaluation studies: the appropriate use of spermatogenic staging. Toxicol. Pathol. 25, 119-131 https://doi.org/10.1177/019262339702500201
  7. Creasy, D. M. 2001. Pathogenesis of male reproductive toxicity. Toxicol. Pathol. 29, 64-76 https://doi.org/10.1080/019262301301418865
  8. Dostal, J. A., C. K. Faber and J. Zandee. 1996. Sperm motion parameters in vas deferens and cauda epididymal rat sperm. Reprod. Toxicol. 10, 231-235 https://doi.org/10.1016/0890-6238(96)00027-5
  9. ICH Harmonized Tripartite Guideline. 1993. Detection of toxicity to reproduction for medicinal products
  10. Ichihara, G., N. Asaeda, T. Kumazawa, T. Tagawa, M. Kamijima, X. Yu, H. Kondo, T. Nakajima, J. Kitoh, I. J. Yu, Y. H. Moon, N. Hisanaga and Y. Takeuchi. 1997. Testicular and haematopoietic toxicity of 2-bromopropane, a substitute for ozone layer-depleting chlorofluorocarbons. J. Occup. Health 39,57-63 https://doi.org/10.1539/joh.39.57
  11. Kim, J. C., H. S. Lee, H. I. Yun and M. K. Chung. 2000. Evaluation of the testicular toxicity caused by 2-bromopropane in rats. Korean J. Vet. Res. 40, 361-371
  12. Kim, J. C., K. H. Lim and M. K. Chung. 1999. Testicular cytotoxicity of DA-125, a new anthracycline anticancer agent, in rats. Reprod. Toxicol. 13, 391-397 https://doi.org/10.1016/S0890-6238(99)00028-3
  13. Kim, J. C., S. H. Kim, D. H. Shin, T. H. Ahn, H. C. Kim, Y. B. Kim, C. Z. Jiang, J. Han and M. K. Chung. 2004a. Effects of prenatal exposure to the environmental pollutant 2-bromopropane on embryo-fetal development in rats. Toxicology 196, 77-86 https://doi.org/10.1016/j.tox.2003.11.006
  14. Kim, J. C., D. H. Shin, S. H. Kim, J. K. Kim, S. C. Park, W. C. Son, H. S. Lee, J. E. Suh, C. Y. Kim, C. S. Ha and M. K. Chung. 2004b. Subacute toxicity study of a new camptothecin anticancer agent CKD-602 administered by intravenous injection to rats. Regul. Toxicol. Pharmacol. 40, 356-369
  15. Matsui, H. and M. Takahashi. 1999. A novel quantitative morphometry of germ cells for the histopathological evaluation of rat testicular toxicity. J. Toxicol. Sci. 24, 17-25 https://doi.org/10.2131/jts.24.17
  16. Omura, M., Y. Romero, M. Zhao and N. Inoue. 1999. Histopathological evidence that spermatogonia are the target cells of 2-bromopropane. Toxicol. Lett. 104, 19-26 https://doi.org/10.1016/S0378-4274(98)00350-6
  17. Riecke, K. and R. Stahimann. 2000. Test systems to identify reproductive toxicants. Andrologia 32, 209-218 https://doi.org/10.1046/j.1439-0272.2000.00422.x
  18. Robb, G. W., R. P. Amann and G. J. Killian. 1978. Daily sperm production and epididymal sperm reserves of pubertal and adult rats. J. Reprod. Fertil. 54, 103-107 https://doi.org/10.1530/jrf.0.0540103
  19. Russel, J. D., R. A. EttIin, A. P. Sinha Hikim and E. D. Clegg. 1990. Histological and Histopathological Evaluation of The Testis. pp. 1-286, Cache River Press, Oearwater, Florida
  20. Sakai, T., M. Takahashi, K. Mitsumori, K. Yasuhara, K. Kawashima, H. Mayahara and Y. Ohno. 2000. Collaborative work to evaluate toxicity on male reproductive organs by repeated dose studies in rats: overview of the studies. J. Toxicol. Sci. 25 (Suppl. 1), 1-21
  21. Seed, J., R. E. Chapin, E. D. Clegg, L. A. Dostal, R. H. Foote, et al. 1996. Methods for assessing sperm motility, morphology, and counts in the rat, rabbit, and dog: a consensus report. Reprod. Toxicol. 10, 237-244 https://doi.org/10.1016/0890-6238(96)00028-7
  22. Sharpe, R. M. 1994. Regulation of spermatogenesis. In: The Physiology of Reproduction. Second Edition. Knobil E and Neil JD (eds), pp. 1363-1434, Raven Press, New York
  23. Son, H. Y, Y. B. Kim, B. H. Kang, S. W. Cho, C. S. Ha and J. K. Roh. 1999. Effects of 2-bromopropane on spermatogenesis in the Sprague-Dawley rat. Reprod. Toxicol. 13, 179-187 https://doi.org/10.1016/S0890-6238(99)00005-2
  24. Son, W. C., J. C. Kim and I. J. Yu. 2003. The recommended approaches for the evaluation of testicular toxicity with awareness of the spermatogenic cycle and quantitative testicular toxicity evaluation methods. J. Toxicol. Pub. Health 19, 83-90
  25. Takayama, S., M. Akaike, K. Kawashima, M. Takahashi and Y. Kurokawa. 1995. A collaborative study in Japan on optimal treatment period and parameters for detection of male fertility disorders induced by drugs in rats. J. Am. Coll. Toxiciol. 14, 266-292 https://doi.org/10.3109/10915819509008702
  26. Takahashi, O. and S. Oishi. 2003. Testicular toxicity of dietarily or parenterally administered bisphenol A in rats and mice. Food Chem. Toxicol. 41, 1035-1044 https://doi.org/10.1016/S0278-6915(03)00031-0
  27. Wu, X., A. S. Faqi, J. Yang, B. P. Pang, X. Ding, X. Jiang and I. Chahoud. 2002. 2-Bromopropane induces DNA damage, impairs functional antioxidant cellular defenses, and enhances the lipid peroxidation process in primary cultures of rat Leydig cells. Reprod. Toxicol. 16, 379-384 https://doi.org/10.1016/S0890-6238(02)00039-4
  28. Zenick, H. and E. Clegg. 1989. Assessment of male reproductive toxicity: a risk assessment approach. In Principles and Methods of Toxicology (Hayes AW, ed), Second Edition, pp. 275-309, Raven Press, New York
  29. Zhao, L. X., E. K. Kim, H. T. Lim, Y. S. Moon, N. H. Kim, T. H. Kim, H Choi, W. Chae, T. C. Jeong and E. S. Lee. 2002. Synthesis, characterization and in vitro identification of $N^{7}$-guanine adduct of 2-bromopropane. Arch. Pharm. Res. 25, 39-44 https://doi.org/10.1007/BF02975258