• Toxicological Research
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• v.18 no.4
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• pp.393-396
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• 2002

This study was carried out to evaluate the ostrogenic/antiandrogenic activity of Puerariae Radix in Sprague-Dawley rats. It has known that diverse phytoestrogen were included in some Puerariae Radix, especially in Pueraria mirifica. The Uterotrophic assay and Hershberger assay were performed to evaluate the ostogenic/antiandrogenic activity of various Puerariae Radix (Pueraria thunbergiana, Pueraria mirifica and Butea superba). In Uterotrophic assay, the extracts of Puerariae Radix were administered subcutaneously to immature female SD rats from 19 to 21 days of age. The wet uterus and vaginal weighs significantly increased in the group only treated with extracts of Pueraria mirifica. But, in Hersh-berger assay, all extracts of Puerariae Radix did not show any effects in the castrated rats. These results suggest that Pueraria mirifica has not undrogenic/antiandrogenic effect but potent estrogenic effect. It is possible that components of Pueraria mirifica may act as endocrine disruptor in human body.

• Biomolecules & Therapeutics
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• v.8 no.2
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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.

• Toxicological Research
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• v.21 no.2
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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.

• Environmental Mutagens and Carcinogens
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• v.20 no.1
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• pp.1-6
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• 2000

자궁내 반응에 대한 용량 의존적인 효과를 검색하기 위해 에스트로겐 agonist 및 antagonist를 사용하여 미성숙 랫드에서 uterotrophic assay를 실시하였다. 김(2000) 등은 이미 난소절제랫드를 이용한 3-day uterotrophic assay에서 에스트로겐성 물질에 대한 검색 시험을 실시한 바 있다. 양성대조물질인 17-ethinyl estradiol (EE)을 미성숙랫드에 매일 3일간 피하 투여하였으며, EE의 에스트로겐 활성을 차단하기 위한 항에스트로겐성 물질인 ZM189.154은 EE와 병행 투여하였다. 본 시험 결과 EE 1.0$\mu\textrm{g}$/kg 이상 투여군에서는 자공 비대와 fluid 저류등이 대조군에 비해 육안적으로 뚜렷하게 나타났다. 자궁무게(wet 및 blotted)는 용량 의존적으로 증가하였으며, 특히, EE 1.0$\mu\textrm{g}$/kg 이상에서는 유의성있는 증가를 나타내었다. 또한 질무게 증가도 자궁무게 증가와 일치 하였다. EE 0.3$\mu\textrm{g}$/kg을 투여한 후 ZM189, 154 0.1 mg/kg 및 1.0 mg/kg 투여와 비교할 때 EE에 의해 유도된 에스트로겐 활성이 용량 의존적으로 감소되었으나 유의성 있는 결과는 ZM189, 154 1.0 mg/kg군에서만 나타났다. 본 연구결과 EE는 uterotrophic assay에서 자궁 무게 (wet와 blotted)를 용량 의존적으로 증가시켰으며, ZM189, 154 1.0mg/kg은 EE의 에스트로겐 활성을 유의하게 감소시켰다. 따라서 미성숙 랫드를 이용한 3-day uterotrophic assay는 내분비계 장애물질 검색 시험법으로 우수하다는 것을 알 수 있었다.

• Korean Journal of Food Science and Technology
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• v.32 no.4
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• pp.964-969
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• 2000

Di-(2-ethylhexyl) adipate(DEHA) has been used extensively as a plasticizer in the manufacture of plastic products such as PVC films. Though, phthalate esters plasticizers have been known to induce endocrine system-mediated responses, few studies have been conducted for the screening of estrogenic activity of DEHA, an adipate plasticizer. This study was initiated to evaluate the estrogenic activity of DEHA by in vitro E-screen assay and in vivo uterotrophic assay. MCF-7 human breast cancer cells were treated with $DEHA(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$, for 144 hr, and cell proliferation was determined by sulforhodamine B(SRB) assay. DEHA dissolved in corn oil was administered subcutaneously to ovariectomized(OVX) female Sprague-Dawley rats at dosage levels of 0, 2, 20 and 200 mg/kg/day for three consecutive days. Rats were sacrificed 24 hr after final treatment and vagina and uterus(wet and blotted) weights were obtained. E-screen assayed DEHA did not generate cell proliferation at treated concentrations$(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$, whereas 17 ${\beta}-estradiol$(E2), the positive control, induced cell proliferation at low concentrations$(5{\times}10^{-14}{\sim}5{\times}10^{-9}\;M)$. In the uterotrophic assay, DEHA did not change vagina and uterus(wet and blotted) weights at dosage levels up to 200 mg/kg/day treatment. These results demonstrated that DEHA did not exhibit the estrogenic activity as determined by in vitro E-screen assay and in vivo uterotrophic assay.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.293-299
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• 2000

Many methods have been developed for screening chemicals with hormonal activity. Using recombinant yeasts expressing either human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127 (YER)] or androgen receptor [S. cerevisiae AR + 8320 (YAR)], we evaluated the hormonal activities of several chemicals by induction of ${\beta}-galactosidase$ activity. The chemicals were $17{\beta}-estradiol$ (E2), testosterone (T), ${\rho}-nonylphenol$ (NP), bisphenol A (BPA), genistein (GEN), 2-bromopropane (2-BP), dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and butylparaben (BP). To assess the estrogenicity of NP, the result of the in vitro recombinant yeast assay was compared with an E-screen assay using MCF-7 human breast cancer cells and an uterotrophid assay using ovariectomized mice. In the YER yeast cells, E2, NP, BPA, GEN, and BP exhibited estrogenicity in a doseresponse manner, while TCDD did not. All the chemicals tested, except T, did not show androgenicity in the YAR yeast cell. The sensitivity of the yeast (YER) assay system to the estrogenic effect of NP was similar to that of the E-screen assay. NP was also estrogenic in the uterotrophic assay. However, in terms of convenience and costs, the yeast assay was superior to the E-screen assay or uterotrophic assay. These results suggest that the recombinant yeast assay can be used as a rapid tool for detecting chemicals with hormonal activities.

• Journal of Environmental Health Sciences
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• v.33 no.5
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• pp.379-385
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• 2007

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.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.11b
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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)

• Biomolecules & Therapeutics
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• v.8 no.1
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• pp.78-83
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• 2000

The objective of this study was to prevalidate the Organization for Economic Cooperation and Development's (OECD) rodent uterotrophic assay as a test method for screening of potential endocrine disrupting chemicals (EDCs). This study was conducted exactly as described in the OECD protocol documents. A positive control substance, 17$\alpha$-ethinyl estradiol (EE), was administered daily for three days to ovariectomized (OVX) Sprague-Dawley rats at various doses for determine the dose-response curve. Additionally, a pure antiestrogenic chemical, ZM189, 154 was administered to OVX rats at the same time EE to determine the effectiveness of the material against blocking the estrogenic effects of EE. At higher concentration of EE (10 $\mu\textrm{g}$/kg), a statistically significant difference in body weight gain and food consumption was observed compared to vehicle controls. In uterine responses, EE produced a dose-related increase in uterus weights compared to vehicle control. These increases were statistically significant at the >1.0 $\mu\textrm{g}$/kg doses. However, a similar dose-response relationship was not observed in vagina weight. A comparison of the two groups receiving ZM189,154 (0.1 and 1.0 mg/kg) with 0.3 $\mu\textrm{g}$/kg of EE and the group receiving only 0.3 $\mu\textrm{g}$/kg of EE showed dose-related decreases in uterus weights. However, statistical significance was shown in 1.0 mg/kg of ZM189,154. In conclusion, administration of EE produced a dose-related increase in uterine (wet and blotted) weights. Additionally, the 1.0mg/kg dose of ZM189,154 was effective in blocking the estrogenic activity of EE. These data suggest 3-day uterotrophic assay using OVX rats may serve as a good tool for EDCs screening.

• Toxicological Research
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• v.16 no.3
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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.