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Assessing Heavy Metals for Estrogenicity Using a Combination of In vitro and In vivo Assays

In vitro 및 In vivo Assay를 통한 중금속의 에스트로겐성 평가

  • Park, Chul (Dept. of Food Science and Nutrition, University of Ulsan) ;
  • Kim, So-Jung (Dept. of Food Science and Nutrition, University of Ulsan) ;
  • Shin, Wan-Chul (Dept. of Food Science and Nutrition, University of Ulsan) ;
  • Kim, Hae-Gyoung (Dept. of Food Science and Nutrition, University of Ulsan) ;
  • Choe, Suck-Young (Dept. of Food Science and Nutrition, University of Ulsan)
  • 박철 (울산대학교 생활과학대학 식품영양전공) ;
  • 김소정 (울산대학교 생활과학대학 식품영양전공) ;
  • 신완철 (울산대학교 생활과학대학 식품영양전공) ;
  • 김혜경 (울산대학교 생활과학대학 식품영양전공) ;
  • 최석영 (울산대학교 생활과학대학 식품영양전공)
  • Published : 2004.11.01

Abstract

The estrogenicities of six heavy metal compounds, which contaminate frequently in foods, were assayed using a combination of in vitro and in vivo assays. The assays were 1) estrogen receptor dependent transcriptional expression assay, 2) E-screen assay and, 3) the uterotropic assay in mice. The chemicals studied were 17$\beta$ -estradiol, diethylstilbestrol (DES), arsenic oxide, bis(tri-n-butyltin), cadmium chloride, chromium chloride, lead acetate, and mercuric chloride. Using the estrogen receptor dependent transcriptional expression assay, the following estrogenicity ranking was measured: bis(tri-n-butyltin) > cadmium chloride > chromium chloride >> mercuric chloride >lead acetate = arsenic oxide. Using E-screen test, the following estrogenicity ranking was measured: bis(tri-n-butyltin) > cadmium chloride > chromium chloride >> mercuric chloride > lead acetate = arsenic oxide. Results from the uterotropic assay showed that bis(tri-n-butyltin), cadmium chloride, chromium chloride caused an increase in uterine wet weight, while lead acetate, mercuric chloride, and arsenic oxide failed to do so. Bis(tri-n-butyltin), cadmium chloride and chromium chloride showed the highest estrogenicity in three assay systems. Recent studies suggesting that bis(tri-n-butyltin), cadmium chloride have estrogenicities are compatible with the present finding. Furthermore, our study is suggesting that chromium chloride may be estrogenic. The results demonstrate that this three level-assay combination (transcriptional activation, cell proliferation, and an in vivo effect in an estrogen-responsive tissue) could serve as a useful method to assess the estrogenicity of heavy metals.

식품오염 관련 중금속들의 에스트로겐성을 in vitro 와 in vivo 분석방법을 병행하여 평가하였다. 분석방법은 1) estrogen receptor dependent transcriptional expression 분석법, 2) E-screen assay 그리고, 3) 마우스 자궁비대시험 (uterotropic assay)을 사용하였다. 시험에 사용한 물질로는 $17\beta$-estradiol, diethylstilbestrol(DES), arsenic oxide, bis (tri-n-butyltin), cadmium chloride, chromium chloride, lead acetate, mercuric chloride을 사용하였다. Estrogen receptor dependent transcriptional expression 분석 결과, bis(tri-nbutyltin) > cadmium chloride > chromium chloride 순으로 에스트로겐성이 크게 나타났으며, mercuric chloride, lead acetate, arsenic oxide는 거의 나타나지 않았다. E-screen test 결과, bis(tri-n-butyltin) > cadmium chloride > chromium chloride 순으로 에스트로겐성이 크게 나타났으며, mercuric chloride, lead acetate, arsenic oxide는 거의 나타나지 않았다. 자궁비대시험 결과도 마찬가지로 bis(tri-nbutyltin), cadmium chloride, chromium chloride은 자궁중량 비대를 크게 초래하였으며, 반면에 mercuric chloride, lead acetate, arsenic oxide는 그러한 효과가 미약하거나 없었다. 세 분석방법 결과 bis(tri-n-butyltin), cadmium chloride, chromium chloride 순으로 에스트로겐성이 크게 나타났다. 이러한 결과는 최근 bis(tri-n-butyltin)과 cadmium chloride이 에스트로겐성이 있다는 다른 연구결과들과 잘 일치하며, 또한 크롬화합물도 에스트로겐성이 있다는 것을 새롭게 제시하고 있다. 본 연구는 세 단계 수준(전사활성화단계, 세포증식작용, in vivo assay)의 분석을 병행함으로써 수많은 중금속의 에스트로겐성을 효과적으로 평가할 수 있다는 것을 제시해주고 있다.

Keywords

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