Immatured Type Uterotrophic Assay for Estrogenicity Evaluation of DEHA

미성숙 랫드 자궁비대반응시험을 이용한 DEHA의 내분비계 장애작용 평가

  • Park, Ki-Dae (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Han, Beom-Seok (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Jeong, Ja-Young (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Oh, Jae-Ho (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Cho, Wan-Seob (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Cho, Min-Jeong (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Choi, Mi-Na (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Kim, Sung-Joon (Korea Food and Drug Administration, National Institute of Toxicological Research) ;
  • Kim, Seung-Hee (Korea Food and Drug Administration, National Institute of Toxicological Research)
  • 박기대 (식품의약품안전청 국립독성과학원) ;
  • 한범석 (식품의약품안전청 국립독성과학원) ;
  • 정자영 (식품의약품안전청 국립독성과학원) ;
  • 오재호 (식품의약품안전청 국립독성과학원) ;
  • 조완섭 (식품의약품안전청 국립독성과학원) ;
  • 조민정 (식품의약품안전청 국립독성과학원) ;
  • 최미나 (식품의약품안전청 국립독성과학원) ;
  • 김성준 (식품의약품안전청 국립독성과학원) ;
  • 김승희 (식품의약품안전청 국립독성과학원)
  • Published : 2007.10.30


This study was aimed to investigate the estrogenic activity of Di-(2-ethylhexyl) adipate (DEHA) using immatured type uterotrophic assay. SD rats were treated with DEHA (40, 200, 1000mg/kg/day), estradiol-3-benzoate (EB) $(1{\mu}g/kg/day)$ as positive control on the assay. In immatured-type uterotrophic assay, relative organ weights of kidney and reproductive organs such as ovary at high-dose group were significantly increased compared to those of vehicle control group. DEHA did not influence the levels of serum FSH and LH, and uterine morphological changes such as luminal epithelial height, myometrial thickness and numbers of uterine gland, and BrdU indices. In these results, there was no significant variation by DEHA treatment, suggesting that DEHA appears not to be a endocrine disrupter with estrogenic activity.


DEHA;immatured type uterotrophic assay;estrogenic activity


  1. 식품의약품안전청 : 내분비계 장애(추정)물질 독성자료집. 141, 1999
  2. Petersen, J. H., Naamansen, E. T. and Nielsen, P. A. : PVC cling film in contact with cheese: health aspects related to global migration and specific migration of DEHA. Food Additives Contaminants, 12(2), 245-253, 1995
  3. Moody, D. E. and Reddy, J. K. : Serum triglyceride and cholesterol contents in male rats receiving diets containing plasticizers and analogues of the ester 2-ethylhexanol. Toxicology Letters, 10(4), 379-383, 1982
  4. Birner, P., Ritzi, M., Musahl, C., Knippers, R., Gerdes, J., Voigtlander, T., Budka, H. and Hainfellner, J. A. : Immunohistochemical detection of cell growth fraction in formalin-fixed and paraffin-embedded murine tissue. The American Journal of Pathology, 158(6), 1991-1996, 2001
  5. Bosq, J. and Bourhis, J. : Bromodeoxyuridine (BrdU). Analysis of cellular proliferation. $Ann{\frac{1}{4}}OE$ of Pathology, 17(3), 171-178, 1997
  6. Shin, J. H., Moon, H. J., Kim, T. S., Kang, I. H., Ki, H. Y., Choi, K. S. and Han, S. Y. : Repeated 28-day oral toxicity study of vinclozolin in rats based on the draft protocol for the 'Enhanced OECD Test Guideline No. 407' to detect endocrine effects. Archives of Toxicology, 80(9), 547-554, 2006
  7. Kim, E. J. : Study on Anti-estrogenic Activity of DEHP as an Endocrine Disruption Chemical. Korean Journal of Environmental Health, 29(2), 7-15, 2003
  8. 환경부 : 유해화학물질관리기본계획(2001-2005). 236, 2000
  9. Ashby, J., Odum, J. and Foster, J. R. : Activity of raloxifene in immature and ovariectomized rat uterotrophic assays. Regulatory Toxicology and Pharmacology, 25(3), 226-231, 1997
  10. Odum, J., Lefevre, P. A., Tittensor, S., Paton, D., Routledge, E. J., Beresford, N. A., Sumpter, J. P. and Ashby, J. : The rodent uterotrophic assay: critical protocol features, studies with nonyl phenols, and comparison with a yeast estrogenicity assay. Regulatory Toxicology and Pharmacology, 25(2), 176-188, 1997
  11. Laurence, C., Angela, J. M. and John, G. : Migration from plasticized films into foods. 4. Use of polymeric plasticizers and lower levels of di-(2-ethylhexyl adipate plasticizers in PVC films to reduce migration into food. Food Additives Contaminants, 15(3), 277-282, 1988
  12. Kaufmann, W., Deckardt, K., McKee, R. H., Butala, J. H. and Bahnemann, R. : Tumor induction in mouse liver: di-isononyl phthalate acts via peroxysome proliferation. Regulatory Toxicology and Pharmacology. 36(2), 175-183, 2002
  13. Singh, A. R., Lawrence, W. H. and Autian, J. : Dominant lethal mutations and antifertility effects of di-2-ethylhexyl adipate and diethyl adipate in male mice. Toxicology and Applied Pharmacology, 32(2), 566-576, 1975
  14. Lake, B. G., Price, R. J., Cumminghame, M. E. and Walters D. G. : Comparison of the effects of di-(2-ethylhexyl) adipate on hepatic peroxisome proliferation and cell replication in the rat and mouse. Toxicology, 123(3), 217-226, 1997
  15. Kluwe, W. M., McConnell, E. E., Huff, J. E., Haseman, J. K., Douglas, J. F. and Hartwell, W. V. : Carcinogenicity testing of phthalate esters and related compounds by the National Toxicology Program and the National Cancer Institute. Environmental Health Perspectives, 45, 129-133, 1982
  16. Kang, K. S., Kim, H. S., Ryu, D. Y., Che, J. H. and Lee, Y. S. : Immature uterotrophic assay is more sensitive than ovariectomized uterotrophic assay for the detection of estrogenicity of p-nonylphenol in Sprague-Dawley rats. Toxicology Letters, 20, 118(1-2), 109-115, 2000
  17. Han, S. Y., Kim, H. S., Han, S. K., Rhee, Y., Yang, K. H. and Park, K. R. : Study on the estrogenic activity of di-(2-ethylhexyl) adipate in E-screen assay and uterotrophic assay. Korean Journal of Food Science Technology, 32(4), 964-969, 2000
  18. Kim, P. G. and Yang, Y. H. : Effects of Butyl Benzyl Phthalate on Dams and F1 during Lactation Period of Rats. Korean Journal of Environmental Health, 29(2), 16-22, 2003
  19. Jordan, V. C. and Gosden, B. : Differential antiestrogen action in the immature rat uterus: a comparison of hydroxylated antiestrogens with high affinity for the estrogen receptor. Journal of Steroid Biochemistry, 19(3), 1249-1258, 1983
  20. Cho, S. D., Kim, J. H., Kim, D. Y., Lee, Y. S. and Kang, K. S. : Pre-validation study for OECD enhanced test guideline 407 protocol by gavage for 4 weeks using propylthiouracil and tamoxifen. Toxicology Letters, 144(2), 195-204, 2003
  21. Melnick, R. L., Morrissey, R. E. and Tomaszewski, K. E. : Studies by the National Toxicology Program on di(2-ethylhexyl)phthalate. Toxicology and Industrial Health, 3(2), 99-118, 1987
  22. Miyagawa, M., Takasawa, H., Sugiyama, A., Inoue, Y., Murata, T., Uno, Y. and Yohikawa, K. : The in vivo-in vitro replicative DNA synthesis (RDS) test with hepatocytes prepared from male B6C3F1 mice as an early prediction assay for putative nongenotoxic (Ames-negative) mouse hepatocarcinogens. Mutation Research, 343, 157-183, 1995
  23. OECD : Validation Protocol for the Uterotrophic Assay. Task Force on Endocrine Disruptor Testing and Assessment. 1999
  24. NTP : Carcinogenesis bioassay of di-2-ethylhexyl) adipate in F344 rats and B6C3F1 mice. National Toxicology Program Technical Report, Series No. 212, 1982
  25. Lee, H. C., Yamanouchi, K. and Nishihara, M. : Effects of Perinatal Exposure to Phthalate/adipate esters on hypothalamic gene expression and sexual behavior in rats. The Journal of Reproduction and Development. 52(3), 343-52, 2006
  26. Peterson, J. H. and Breindahl, T. : Specific migration of di-(2-ethylhexyl)adipate (DEHA) from plasticized PVC film: results from an enforcement campaign. Food Additives Contaminants, 15(5), 600-608, 1998
  27. EPA EDSTAC : Endocrine Disrupotr Screening and Testing Advisory Committee Final Report, 1998
  28. Choi, K., Hwang, S. D., Kwon, E. and Kim, P. G. : The Reproductive Toxicity by Combined Treatment of Bisphenol A and Butyl Benzyl Phthalate During Gestation, Lactation Period in Rats. Korean Journal of Environmental Health, 30(2), 71-78, 2004