The Estrogenic Effects of Phthalates(DEHP, DBP) in Yeast Recombinant Assay

효모재조합 검색시험법을 이용한 DEHP, DBP의 에스트로젠 효과

  • Jung, Ji-Youn (Department of Companion and Laboratory Animal Science, Kongju National University)
  • Published : 2007.09.30

Abstract

Di(2-ethylhexyl) phthalate (DEHP) and di-n-butyl phthalate (DBP) were screened for estrogenic activity using a recombinant yeast screening system consisted with estrogen receptors and ${\beta}-galactosidase$ as reporter gene. The chemicals showed estrogenic activity in ranges of $1{\times}10^{-10}\;to\;1{\times}10^{-7}M(DEHP)\;and\;of\;1{\times}10^{-9}M\;to\;1{\times}10^{-6}M(DBP)$ respectively. $17{\beta}-estradiol$, as a positive control of, showed maximal activity at $1{\times}10^{-9}M$. The concentration of half-maximal estrogenic activity was $1{\times}10^{-9}M$ for both chemicals. However, the concentration of maximal estrogenic activity was $1{\times}10^{-7}M$ for DEHP and $1{\times}10^{-8}M$ M for DBP. These results suggested that DBP was higher in relative potencies and more sensitive than DEHP. In conclusion, DEHP and DBP were both estrogenic, even though DBP was more reactive to estrogen receptor.

에스트로젠 수용체와 리포터 유전자인 ${\beta}-galactosidase$가 도입된 효모재조합검색시험법을 이용하여 DEHP와 DBP의 내분비계 장애작용을 검색하였다. 양성대조 시험물질로 $17{\beta}-estradiol$을 설정하여 DEHP와 DBP의 에스트로젠 활성을 비교분석 하였을 때, $17{\beta}-estradiol$의 경우 $10^{-9}M$에서 가장 활성이 높게 관찰되었다. DEHP의 경우 $10^{-10}M$에서 $10^{-7}M$까지 시험하였을 때 농도의존적으로 에스트로젠활성이 증가하였으며, $10^{-7}M$의 경우 가장 강력한 에스트로젠활성을 보였다. DBP의 경우 $10^{-9}M$에서 $10^{-6}M$까지 에스트로젠활성이 관찰되었다. DEHP와 DBP의 경우 최대활성화 대비 50% 이상의 활성도를 보이기 시작하는 농도가 $10^{-9}M$로 나타나서 비슷한 농도에서 에스트로젠 활성화가 이루어지는 것으로 추측할 수 있었다. 그러나, 에스트로젠 최대활성화 시의 농도를 비교해 보면 DBP가 DEHP보다 10배 낮은 농도에서 최대활성치가 관찰되었기 때문에 DBP가 DEHP보다 에스트로젠 작용에 더 민감하게 반응하는 것으로 판단할 수 있었다. 결과적으로 본 실험에 사용되어진 시험물질인 DEHP와 DBP는 효모재조합시험법에 있어서 에스트로젠 활성을 유도하는 것으로 판단되어지며 감수성에 있어서는 DBP가 DEHP보다 높은 것으로 여겨진다.

Keywords

References

  1. Carlsen, E., Giwercman, A., Keiding, N., and Skakkebaek, N.E.: Evidence for decreasing quality of semen during past 50 years. BMJ. 305, 609-613 (1992) https://doi.org/10.1136/bmj.305.6854.609
  2. Sharpe, R.M. and Skakkebaek, N.E.: Are oestrogens involved in falling sperm counts and disorders of the male reproductive tract? Lancet. 341, 1392-1395 (1993) https://doi.org/10.1016/0140-6736(93)90953-E
  3. Soto, A.M., Justicia, H., and Wray, J.W., and Sonnenschein C.: p-Nonyl-phenol: An estrogenic xenobiotic released from 'modified' polystyrene. Environ Health Perspect. 92, 167-173 (1991) https://doi.org/10.2307/3431154
  4. White, R., Jobling, S., Hoare, S.A., Sumpter, J.P., and Parker, M.G..: Environmentally persistent alkylphenolic compounds are estrogenic. Endocrinology. 135, 175-182 (1994) https://doi.org/10.1210/en.135.1.175
  5. Jobling, S., Reynolds, T., White, R., Parker, M.G., and Sumpter, J.P.: A variety of environmentally persistent chemicals, including some phthalate plasticizers, are weakly estrogenic. Environ Health Perspect. 103, 582-587 (1995) https://doi.org/10.2307/3432434
  6. Sonnenschein, C., Soto, A.M., Fernandez, M.F., Olea, N., Olea-Serrano, M.F., and Ruiz-Lopez, M.D.: Development of a marker of estrogenic exposure in human serum. Clin Chem. 41, 1888-1895 (1995)
  7. Soto, A.M., Sonnenschein, C., Chung, K.L., Fernandez, M.F., Olea, N., and Serrano, F.O.: The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants. Environ Health Perspect. 103, 113-122 (1995) https://doi.org/10.1289/ehp.95103s7113
  8. Sharpe, R.M., Fisher, J.S., Millar, M.M., Jobling, S., and Sumpter, J.P.: Gestational and lactational exposure of rats to xenoestrogens results in reduced testicular size and sperm production. Environ Health Perspect. 103, 1136-1143 (1995) https://doi.org/10.2307/3432610
  9. Wine, R.N., Li, L.H., Barnes, L.H., Gulati, D.K., and Chapin, R.E.: Reproductive toxicity of di-n-butylphthalate in a continuous breeding protocol in Sprague-Dawley rats. Environ Health Perspect. 105, 102-107 (1997) https://doi.org/10.2307/3433070
  10. Jaeger, R.J. and Rubin, R.J.: Migration of a phthalate ester plasticizer from polyvinyl chloride blood bags into stored human blood and its localization in human tissues. N Engl J Med. 287, 1114-1118 (1972) https://doi.org/10.1056/NEJM197211302872203
  11. Giam, C.S., Chan, H.S., Neff, G.S., and Atlas, E.L.: Phthalate ester plasticizers: a new class of marine pollutant. Science. 199, 419-421 (1978) https://doi.org/10.1126/science.199.4327.419-a
  12. Sharman, M., Read, W.A., Castle, L., and Gilbert, J.: Levels of di-(2-ethylhexyl)phthalate and total phthalate esters in milk, cream, butter and cheese. Food Addit Contam. 11, 375-385 (1994) https://doi.org/10.1080/02652039409374236
  13. Shibko, S.I. and Blumenthal, H.: Toxicology of phthalic acid esters used in food-packaging material. Environ Health Perspect. 3, 131-137 (1973) https://doi.org/10.2307/3428039
  14. Dirven, H.A., van den Broek, P.H., Arends, A.M., Nordkamp, H.H., de Lepper, A.J., Henderson, P.T., and Jongeneelen, F.J.: Metabolites of the plasticizer di(2-ethylhexyl)phthalate in urine samples of workers in polyvinylchloride processing industries. Int Arch Occup Environ Health. 64, 549-554 (1993) https://doi.org/10.1007/BF00517699
  15. Ahel, M., McEvoy, J., and Giger, W.: Bioaccumulation of the lipophilic metabolites of nonionic surfactants in freshwater organisms. Environ Pollut. 79, 243-248 (1993) https://doi.org/10.1016/0269-7491(93)90096-7
  16. Albro, P.W. and Lavenhar, S.R.: Metabolism of di(2-ethylhexyl) phthalate. Drug Metab Rev. 21, 13-34 (1989) https://doi.org/10.3109/03602538909029953
  17. Gray, T.J. and Gangolli, S.D.: Aspects of the testicular toxicity of phthalate esters. Environ Health Perspect. 65, 229-235 (1986) https://doi.org/10.2307/3430187
  18. Siddiqui, A and Srivastava, S.P.: Effect of di(2-ethyl hexyl)phthalate administration on rat sperm count and on sperm metabolic enzymes. Bull Environ Contam Toxicol. 48, 115-119 (1992)
  19. Davis, B.J., Maronpot, R.R, and Heindel, J.J.: Di-(2-ethylhexyl) phthalate suppresses estradiol and ovulation in cycling rats. Toxicol Appl Pharmacol. 128, 216-223 (1994) https://doi.org/10.1006/taap.1994.1200
  20. Gangolli, S.D.: Testicular effects of phthalate esters. Environ Health Perspect. 45, 77-84 (1982) https://doi.org/10.2307/3429387
  21. Ousterhout, J., Struck, R.F., and Nelson, J.A.: Estrogenic activities on methoxychlor metabolites. Biochem Pharmacol. 30, 2869-2871 (1981) https://doi.org/10.1016/0006-2952(81)90429-9
  22. Ashby, J., Lefevre, P.A., Odum, J., Harris, C.A., Routledge, E.J., and Sumpter, J.P.: Synergy between synthetic oestrogens? Nature. 385, 494-494 (1997)