• Biomolecules & Therapeutics
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• 제8권2호
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• pp.147-152
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• 2000

The rodent uterotrophic assay is currently recommended as one of the primary in vivo assays far endocrine disrupting chemicals by the Organization for Economic Cooperation and Development (OECD) and Endocrine Disruptor Screening and Testing Advisory Committee (US EPA EDSTAC). Generally, this assay relies on the rapid increase in uterus and vagina weights when exposed to estrogenic compounds. Phthalate esters have been used extensively as a plasticizer in the manufacture of plastic products such as PVC films and medical devices. Recently, phthalate esters have been shown to induce endocrine system mediated responses. However, a flew studies have been conducted for the screening of their estrogenic activity. In this study the estrogenic activity of seven phthalate esters, butyl benzyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP), di-n-butylphthalate (DBP), diethylphthalate (DEP), di-n-pentylphthalate (DPF), di-n-propylphthalate (DPrP) and dicyclohexylphthalate (DCHP), was examined in uterotrophic assay. Phthalate esters dissolved in corn oil were administered to ovariectomized (OVX) female Sprague-Dawley rats by sub-cutaneous injection for three consecutive days. fiats were sacrificed 24h after final treatment, and then uterus and vagina weights were deter mined. All phthalate esters tested in this assay did not change talc uterus and vagina weights at dosage levels up to 200 mg/kg/day treatment. These results demonstrated that phthalate esters did not exhibit estrogenic activity in vivo uterotrophic assay.

• 한국식품과학회지
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• 제32권4호
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• pp.964-969
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• 2000

랩 등 PVC 제품의 가소제인 DEHA의 에스트로겐 활성을 검색하기 위하여 내분비계 장애작용 중 에스트로겐성 작용을 검색하는 대표적인 방법인 in vitro E-screen assay와 OECD 및 미국 EPA EDSTAC에서 권고하고 있는 in vivo 난소절제 랫드를 이용한 자궁 비대반응시험(uterotrophic assay)을 실시하였다. 시험결과 $DEHA(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$는 MCF-7세포의 증식을 유발하지 않았고 난소절제 랫드에 200 mg/kg/day 용량까지 투여하여도 자궁 및 질 무게에 영향을 미치지 않았다. 따라서 본 실험결과 DEHA는 E-screen assay 및 자궁비대반응시험에서 에스트로겐성 작용이 없는 것으로 확인되었으나, 이들 물질의 에스트로겐성에 대한 결론을 내리기 위해서는 여러 종류의 in vitro 시험 data 및 in vivo 시험 data가 더 보충되어야 할 것으로 생각된다.

• Toxicological Research
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• 제21권2호
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• pp.175-178
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• 2005

Butyl $\rho$-hydroxybenzoic acid (butyl paraben, BP) is a homologous series of parabens and is widely used as a preservative in cosmetic and pharmaceutical products. The purpose of this study was to investigate the estrogenic/antiandrogenic activities of BP in animals. For that, we performed an uterotrophic assay and a Hershberger assay in rats. In uterotrophic assay, BP was administered subcutaneously to immature female SD rats (18 days old) for 3 consecutive days. The wet and dry uterus weights were significantly increased in the groups treated with BP in dose­dependent manner. In case of Hershberger assay, BP significantly reduced the weight of seminal vesicle of castrated rats. And other accessory organ/glands - prostate, Cowper's glands, bulbocavernosus muscle and glans penis were also slightly decreased. The results of this study suggested that BP showed estrogenic and anti-androgenic activities in vivo.

• Molecular & Cellular Toxicology
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• 제2권2호
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• pp.106-113
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• 2006

Phthalate analogues are a plasticizer and solvent used in industry. Phthalates were classified in the category of "suspected" endocrine disruptors. The purpose of our study was to screen and elucidate the endocrine disrupting activity of seven phthalate analogues. E-screen assay was performed in MCF-7 human breast cancer cells with seven phthalate analogues. In this cell proliferation assay, benzyl butyl phthalate (BBP) and dibutyl phthalate (DBP) showed high estrogenic activity. Their relative proliferation efficiencies (RPE) were 109 and 106%, respectively. In vitro estrogen receptor (ER) binding assay, BBP, di-n-octyl phthalate (DOP) and dinonyl phthalate (DNP) showed weak relative binding affinity (RBA: 0.02%) compared to $17{\beta}-estradiol\;(E2)$ (RBA: 100%). In uterotrophic assay, E2 produced a significant increase, whereas four tested phthalate analogues had potential estrogenic effects in vitro did not increased in uterus weight in immature rats. From these results, we demonstrated that phthalate analogues exhibit weak estrogenic activity in vitro assays at high concentrations. Although phthalates induced an increase in MCF-7 cell proliferation by an estrogenic effect, they could not induce a uterus weight increase in vivo. From these, we may suggest that these phthalate analogues are easily metabolized to inactive forms in vivo. Further investigation in other in vitro and in vivo experimental systems might be required.

• Toxicological Research
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• 제16권2호
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• pp.109-116
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• 2000

The purposes of our study were to optimize the conditions of the screening and testing methods for endocrine disruptors, to characterize these assays using several compounds with well-defined endocrine activity, and to compare the sensitivity between these assays currently undergoing validation. Two in vitro test systems, MCF-7 cells proliferation (E-screen assay) and competitive binding to estrogen receptors (ER) were selected to evaluate the estrogenic effects. 17$\beta$-Estradiol (E2) and diethylstilbestrol (DES) were used as a positive control in vitro test. Also, E2 and ethinyl estradiol (EE) were used as a positive control in vivo uterotrophic assay. In in vitro test, E2 and DES showed a strong estrogenic response at concentration of 1.0 nM. In uterotrophic assay, E2 (0.3 $\mu\textrm{g}$/kg) and EE (0.3 $\mu\textrm{g}$/kg) produced a significant increase in uterus and vagina weight in both immature and ovariectomized rats. Although we did not com-pared the specificity between in vivo and in vitro assays, these assay systems may serve as a good tool for endocrine disruptors screening methods. Our data indicate that these assay systems exhibit some difference in their sensitivity to the same estrogenic compounds. Therefore, as a first rapid screening assay for estrogenic activity qf unknown chemicals, at least two assay systems should probably be carried out with a view of high sensitivity and standardization conditions. Also, a careful validation tests are necessary to obtain a reasonable degree of reproducibility.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.166-166
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• 2002

In the present study, estrogenic or antiestrogenic activity of cypermethrin, a pyrethroid insecticide was investigated. We used immature rat uterotrophic assay, estrogen-responsive calbindin-D9k (CaBP-9k) gene expression assay and luciferase reporter gene assay for measure of estrogenic potential of cypermethrin.(omitted)

• Toxicological Research
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• 제16권3호
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• pp.201-209
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• 2000

In association with the international validation program to establish a rodent uterotrophic assay, we conducted preliminary uterotrophic assay proposed by GECD using immature female rats. In the present study, oral and subcutaneous routes were chosen to compare the effects of estrogenic com-pounds in the two dosing regimens. The reference compound ethinyl estradiol (EE) and the antagonist ZM189154(ZM) were administered by gavage or subcutaneously (s.c.) to immature female SD rats from 20 to 22 days of age. For each study, sixty-six female rats were randomly assigned to eleven groups: Untreated control, EE 0,0.01, 0.03, 0.1, 0.3, 1.0,3.0 and 10.0 $\mu\textrm{g}$/kg, EE 3.0 $\mu\textrm{g}$/kg(gavage)/0.3 $\mu\textrm{g}$/kg(s.c) & ZM 0.1 mg/kg, and EE 3.0 $\mu\textrm{g}$/kg(gavage)/0.3 $\mu\textrm{g}$/kg (s.c) & ZM 1.0 mg/kg. There were no treatment-related changes in clinical signs, body weights, food consumption, and necropsy findings in any groups of two studies. The wet and blotted uterus weights increased dose-dependently. Histopathological examination revealed that diameter of uterine duct, height of uterine luminal epithelium. and height oj vaginal epithelium increased dose-dependently. The proliferating cell nuclear antigen (PCNA) immunoreactive cells were increased in number dose-dependently. The estrogenic effects observed in the present studies occurred at $\geq$ 0.3 $\mu\textrm{g}$/kg of oral dose and $\geq$ 0.1 $\mu\textrm{g}$/kg of s.c. dose. An antagonistic effect of ZM against EE was found in both uterus weight and histopathological parameters. From the results obtained, it can be concluded that dose-dependence of the uterotrophic assay using EE and ZM was well demonstrated by gavage and subcutaneous administration and that the estrogenic effects of EE by s.c. dose were higher than those by gavage administration. In addition, blotted uterus weight was more sensitive than wet uterus weight and vaginal epithelial height was found to be the most sensitive parameter among the parameters examined.

• 한국식품조리과학회지
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• 제21권6호통권90호
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• pp.769-775
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• 2005

미성숙 흰쥐에서 누에 번데기 수용성과 지용성 추출 물 및 한약재가 첨가된 누에 번데기 수용성 추출물을 투여하여 에스트로젠 증식 관련 인자의 발현을 확인한 결과는 다음과 같다. 미성숙 흰쥐에서 누에 번데기 수용성 추출물(KW), 누에 번데기 지용성 추출물(KO) 및 KW에 하수오, 인삼, 울금을 7:1:1:1의 비율로 첨가한 복합체(MK)를 각각 100 mg/kg와 500 mg/kg의 농도로 30일간 경구투여한 결과 체중은 유의적인 변화를 나타내지 않았다. 체중에 대한 자궁의 무게는 정상 대조군에 대하여 실험군에서 KW500을 제외하고는 모두 유의적인 증가를 나타내었으며 K0500, KW100, MK100 순으로 각각 $0.49\%,\;0.48\%,\;0.44\%$의 높은 값을 나타내었다. 체중에 대한 난소의 무게는 정상 대조군에 대해 MK군을 제외하고 모두 유의적인 변화를 보였으며 KW100군이 가장 높은 값을 나타내었다. 미성숙 흰쥐의 혈청 내 AST와 ALT 활성 및 creatinine 농도는 모두 군 간에 유의적인 변화는 보이지 않았다. 즉 모든시료가 간과 신장에 독성을 나타내지 않은 결과이다. $ER\alpha$와 $ER\beta$의 발현을 densitometer로 수치화하여 정상대조군을 기준으로 발현의 증가와 억제를 백분율로 표시한 결과 KW100(100 mg/kg)와 MK500(500 mg/kg) 군이 $ER\alpha$의 발현을 유의적으로 증가시켰으며 KW100(100 mg/kg), KW500(500 mg/kg), 및 MK100(100 mg/kg)군이 $ER\beta$의 발현을 유의적으로 증가시켰다. 따라서 누에 번데기 수용성 추출물 및 한약재(인삼, 하수오, 울금)를 첨가한 누에 번데기 수용성 추출물이 생체에서 높은 에스트로젠 활성을 가지고 있는 것으로 사료된다.

• Journal of Ginseng Research
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• 제47권3호
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• pp.385-389
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• 2023

Background: Ginseng has been used as a traditional medicine for treatment of many diseases and for general health maintenance. Previously, we showed that ginseng did not demonstrate estrogenic property in ovariectomized mouse model. However, it is still possible that disruption of steroidogenesis leading to indirect hormonal activity. Methods: The hormonal activities were examined in compliance with OECD guidelines for detecting endocrine disrupting chemicals: test guideline (TG) No. 456 (an in vitro assay method for detecting steroidogenesis property) and TG No. 440 (an in vivo short-term screening method for chemicals with uterotrophic property). Results: Korean Red Ginseng (KRG) and ginsenosides Rb1, Rg1, and Rg3 did not interfere with estrogen and testosterone hormone synthesis as examined in H295 cells according to TG 456. KRG treatment to ovariectomized mice did not show a significant change in uterine weight. In addition, serum estrogen and testosterone levels were not change by KRG intake. Conclusion: These results clearly demonstrate that there is no steroidogenic activity associated with KRG and no disruption of the hypothalamic-pituitary-gonadal axis by KRG. Additional tests will be performed in pursuit of cellular molecular targets of ginseng to manifest mode of action.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.166-166
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• 2002