본 연구는 자외선 광분해에 의한 비스페놀 A (BPA)의 에 스트로겐 활성 저감에 미치는 하수처리장 방류수 유기물질 의 영향을 조사하였다. 방류수 유기물질과 표준으로 사용한 스와니강 자연 유기물질은 극성에 따라 소수성, 반친수성, 친수성 분획으로 분리하였다. 특이 자외선 흡수 (SUVA) 분 석 결과, 방류수 유기물질은 높는 소수성을 가지고 있는 자 연 유기물질과 다르게 소수성이 낮은 미생물 기원 유기물 질과 유사한 특성을 나타내었다. 3시간의 자외선 조사는 방 류수 및 자연 유기물질의 극성에 따라 SUVA 값을 유의하 게 감소시켰다 (p<0.0001). 유기물질이 없는 조건에서, BPA (5.0×10-5 M)의 상대 에스트로겐 활성도는 자외선 광분해 에 의해 86%에서 63%로 감소하였다. 그러나 유기물질이 있 는 조건에서 상대 에스트로겐 활성도는 평균적으로 68%에 서 37%로 감소하였으며, 유기물질의 종류 (방류수 또는 자연 유기물질) 및 극성 (소수성, 반친수성, 친수성)과 유의한 차이 를 나타내지 않았다 (p>0.05). 결과적으로, 유기물질이 있고 없는 조건에서 자외선 광분해에 의해 감소한 BPA의 상대 에스트로겐 활성도는 각각 31%와 23%였으며, 이것은 방류 수와 자연 유기물질 모두 광분해에 의한 BPA의 에스트로겐 활성 저감을 촉진시킨다는 것을 제시한다.

Endocrine disruptors bind to hormone receptors on sperm membrane, therefore spermatozoa are potentially a useful model for examining estrogenic activities of endocrine disruptors. The objective of this study was to compare the effects of two xenoestrogenic compounds [genistein (Gen) and 4-tert-octylphenol (OP)] to those of two steroids [estrogen (E2) and progesterone (P4)] on boar sperm % motility and motion kinematics of in vitro. Porcine spermatozoa were incubated with various concentrations (0.001～100 μM) of each chemical for 15 or 30 min, and then assessed % motility and sperm motion kinematics using computer assisted sperm analyzer (CASA). Each chemical decreased sperm % motility, and OP decreased VSL and VAP compared with untreated control(p<0.05). E2 stimulated the motion kinematic changes except VCL. Moreover, Gen had effects on VCL and VAP alterations after 30 min incubation. In summary, since all chemicals studied effectively altered sperm % motility and motion kinematics, it was concluded that porcine spermatozoa could be a useful model for in vitro screening of potential endocrine disruptors.

The aim of this study is to evaluate the estrogenic activity of Cucurbita pepo seed extract which includes β-sitosterol and other phytosterols. Sample was extracted from Cucurbita pepo seed by supercritical carbondioxide method and resuspended with ethanol. Estrogenic activity was measured by recombinant yeast assay which detects estrogenic activity using recombinant yeast with high level of estrogenic receptor. However, estrogenic activity of pumpkin seed extract was not found in this study. Based on this data, pumpkin seed extract will not cause estrogenic disturbance.

Background & Objectives : Nonylphenol (NP), a member of alkylphenols, has been widely used in manufacturing antioxidants, The present study was undertaken to examine whether short-term exposure to NP can alter the onset of puberty and associated reproductive parameters such as hormone receptor expressions in prepubertal female rats. Methods : NP (0.1, 1, 10 mg/kg BW) was administered orally at 10:00 from postnatal day 25 (PND 25) through the PND 35. The control group was administered orally each day 200 uL of the sesame oil. At PND 36, all groups were sacrificed at 18:00. The first vaginal opening (V.O.) day was monitored, and the weights of reproductive tissues were measured. Ovaries and uteri were used for analysis of mRNA levels, protein levels and histology. The transcriptional activities of the ovaries (StAR, P405scc, 3β-HSD, 17β-HSD, Aromatase and LH-R) and uteri(ER) were measured using quantitative RT-PCRs. The uterine proteins were used for the ER western blotting. Histological analysis of the reproductive organs was carried out. Results: Administration of NP induced early V.O. when compared to control. The ovarian weights of 1 mg group (0.164±0.010 mg/g BW) was significantly higher than those of control group (0.116±0.007 mg/g BW). Among the steroidogenesis-related genes, mRNA levels of the ovarian P450scc, 3β-HSD and Aromatase are increased significantly in all NP-treated groups. The mRNA levels of StAR and 17β- HSD were unchanged. NP administration (1 mg group) significantly increased the LH-R transcript level. Among the steroid hormone receptor genes, the mRNA level of ER-alpha was significantly elevated in 1mg NP group, while the mRNA levels of ER-beta and PR were unchanged in all NP-treated groups. Interestingly, wetern blotting data for uterine steroid hormone receptors revealed the complete disagreement with the transcriptional activities. Histological status of ovaries and uteri also exerted the occureence of advanced maturation in NP-treated groups. Conclusion: The present study demonstartes that the short-term administration of NP accelerates V.O. and maturation of reproductive organs such as ovary and uterus.

Harmonized actions of ovarian estrogen (E2) and progesterone (P4) regulate cell proliferation and differentiation in the uterus with a spatiotemporal manner. Imbalance between the actions and levels of two major regulators often lead to infertility and gynecological diseases, such as endometriosis and endometrial cancer. While numerous works have shown that reduced expression and/or deletion of uterine factors associated with P4 signaling could disturb uterine physiology, local factor(s) to mediate E2 actions has not been extensively studied yet. Here we demonstrate that early growth response 1 (Egr1), a transcription factor which is rapidly induced in the uterus by E2, is required to maintain coordinated actions of E2 and P4 for uterine receptivity for embryo implantation. Given exogenous gonadotrophins to overcome LHβ deficiency in the pituitary of Egr1(－/－) mice, ovulation, fertilization and embryo development normally occurred in these mice. However, they showed complete failure of embryo implantation with reduced uterine responses to artificial decidualization stimuli. While serum levels of E2 and P4 in Egr1(－/－) mice were comparable, genes regulated by E2 and/or P4 in uterine epithelial cells (ECs) were aberrantly expressed on day 4 of pregnancy. Impaired P4 signaling along with absence of PR in ECs caused hypersensitive E2 responses shown as enhanced expression of E2-responsive genes such Muc1 and Ltf as well as reduced levels of P4-dependent genes, such as Ihh and Areg, in ECs of Egr1(－/－) mice. This is consistent with persistent proliferation in ECs and severely impaired proliferation in stromal cells (SCs) in Egr1(－/－) mice treated with E2+P4. Furthermore, primary co-culture of Egr1(－/－) ECs with Egr1(+/+) SCs and vice versa supported a notion that Egr1 itself is required for proper responses to two major regulators, E2 and P4, in both uterine cell compartments. Collectively, our results show that E2-induced Egr1 participates in P4-dependent modulation on E2 activities in the uterus by regulating a spectrum of genes essential for uterine receptivity and embryo implantation.

Estrogens are ubiquitous signaling molecules that influence nearly every cell type, and exert profound effects on embryonic development, and differentiation. Wnt pathway, which recruits β-catenin into nuclei, and activates The Wnt-dependent transcription factors, also plays an important role in embryonic development and stem cell maintenance, and differentiation. Accumulating evidences indicate that potential convergence between these two pathways in carcinoma cells. However, physiological roles of estrogens in development and differentiation of human embryonic stem cells (hESCs) are relatively unknown. Here, we demonstrated that estrogenic compounds 17α-ethinylestradiol (EE2) and genistein (GEN) significantly increased β-catenin expression in undifferentiated hESCs cultured in feeder-free media. Interestingly, GEN treatement induced an increased trend of mesendodermal gene expressions, and significantly inhibited ectodermal gene expressions (Nestin and Pax6) in embrioid body (EB). Expectantly, GEN increased epithelial-mesenchymal transition (EMT) related gene expression (Snail2, and Twist), whereas decreased E-cadherin on day 6 of EB development. Taken together, these suggest that estrogens may in part the powerful effects on normal hESC differentiation. Mechanistic studies of estrogen signaling continue to suggest novel drug targets for stem cells and will also improve screening methods of developmental toxicity.

본 연구는 내분비 장애물질의 검출을 위하여 간세포의 단층 형성, 생존 및 기능에 미치는 어류 혈청의 영향에 대해 검토하였다. 한국산 메기의 간세포는 자신의 혈청 및 뱀장어, 틸라피아 등 타어종의 혈청에 의해 부착 및 단층이 형성되었으나, FBS는 메기 간세포의 단층을 형성시키지 못했다. 0.5에서 3%의 어류 혈청으로 메기 간세포의 단층을 형성 시킬 수 있는데, 이것은 FBS(5~20%) 사용의 1/10 이하로 적은 양이며, 어류 혈청이 FBS를 대체할

In this study, sixty chemicals including 47 pesticides were screened for estrogenic activity using E-screen assay. MCF-7 cell, used in E-screen assay, is known to be proliferated by addition of estrogenic chemicals. Eight of the measured pesticides showed estrogenic activity at the concentration range of 100-10,000nM. Their relative proliferative effect (RPE) and the relative proliferative potency (RPP) were 20-65% and 0.01-1.0%, respectively, when compared with 1.0nM of β-Estradiol-17-acetate(E2). DDVP and diazinone showed most strong estrogenic potency(RPP; 1.0%) and effect(RPE; 65%) of the eight pesticides. These results are in agreement with estrogenic activity of bisphenol A is known as a positive endocrine disrupter. Also, in this study, paraquat, DDVP, 4- chloro-3-methylphenol, 2,4,6-trichlorophenol and diazinon of the measured pesticides are estimated to estrogenic chemical.