In vitro vitellogenin production by carp (Cyprinus carpio) hepatocytes as a screening method for determining (anti)estrogenic activity of xenobiotics.
In vitro vitellogenin production by carp (Cyprinus carpio) hepatocytes as a screening method for determining (anti)estrogenic activity of xenobiotics.
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鲤鱼（Cyprinus carpio）肝细胞体外产生卵黄蛋白原，作为测定异生物质（抗）雌激素活性的筛选方法。
DOI：
10.1006/taap.1999.8663
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发表时间：
1999
影响因子：
3.8
通讯作者：
vandenBerg,M
中科院分区：
文献类型：
--
作者：
Smeets,JM;Rankouhi,TR;Nichols,KM;Komen,H;Kaminski,NE;Giesy,JP;vandenBerg,M
卵黄蛋白前体卵黄原蛋白（Vtg）是雌鱼和雄鱼肝脏分泌的一种雌激素样物质。在这项研究中，anin vitroassay开发用于测量Vtg诱导，使用培养的原代肝细胞从遗传上一致的品系的鲤鱼（鲤鱼）。Vtg的生产测定间接竞争ELISA法，使用多克隆抗血清对金鱼Vtg，与鲤鱼Vtg交叉反应。在两种性别的肝细胞中，17β-雌二醇（E2）可剂量依赖性地诱导Vtg。E2对Vtg诱导的最低观察效应浓度（LOEC）为2 nM，EC 50为50 - 150 nM，最大响应为2 μM。50和100 μM增塑剂和异雌激素双酚A诱导两种性别肝细胞分泌Vtg。这种鲤鱼肝细胞（CARP-HEP）测定也可用于检测抗雌激素活性，其被测量为E2刺激的Vtg合成的减少。两个众所周知的抗雌激素化合物，他莫昔芬和2，3，7，8-四氯二苯并-p-二恶英（TCDD），进行了测试。TCDD在浓度<0.1 nM时导致雌性肝细胞中Vtg合成减少，使其效力比他莫昔芬高约10，000倍。鲤鱼肝细胞对细胞色素P4501 A（CYP 1A）活性的诱导也很敏感，测定为乙氧基试卤灵O-脱乙基酶（EROD）。根据暴露时间，18或96小时，EROD EC 50值TCDD分别为27或6 pM。使用96孔板形式的CARP-HEP测定提供了筛选大量化合物的（抗）雌激素性质的良好可能性。此外，它可以同时确定芳香烃受体激动剂的特性，测量为CYP 1A诱导。
The yolk protein precursor vitellogenin (Vtg) is secreted by the liver of female as well as male fish, in response to estrogenic compounds. In this study, anin vitroassay was developed for measuring Vtg induction, using cultured primary hepatocytes from genetically uniform strains of carp (Cyprinus carpio). Vtg production was measured by indirect competitive ELISA, using a polyclonal antiserum against goldfish Vtg that cross-reacts with carp Vtg. Vtg was dose-dependently induced by 17β-estradiol (E2) in hepatocytes of both sexes. E2 had a lowest observed effect concentration (LOEC) for Vtg induction of 2 nM, an EC50 between 50 and 150 nM, and a maximum response at 2 μM. The plasticizer and xenoestrogen bisphenol-A induced Vtg secretion by hepatocytes of both sexes at 50 and 100 μM. This carp hepatocyte (CARP–HEP) assay can also be used to detect antiestrogenic activity, which was measured as the reduction of E2-stimulated Vtg synthesis. Two well-known antiestrogenic compounds, tamoxifen and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), were tested. TCDD caused a reduction in Vtg synthesis in female hepatocytes at concentrations <0.1 nM, making it approximately 10,000-fold more potent than tamoxifen. Carp hepatocytes were also sensitive to induction of cytochrome P4501A (CYP1A) activity, measured as ethoxyresorufinO-deethylase (EROD). Depending on the exposure time, 18 or 96 h, EROD EC50 values for TCDD were 27 or 6 pM, respectively. The CARP–HEP assay, using the 96-well plate format, offers good possibilities to screen large numbers of compounds for (anti)estrogenic properties. In addition, it can simultaneously determine aryl hydrocarbon receptor agonist properties, measured as CYP1A induction.
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DOI：
10.1080/15287399409531856
发表时间：
1994-04
期刊：
Journal of toxicology and environmental health
影响因子：
--
作者：
D. Spink;J. Johnson;S. Connor;K. Aldous;J. Gierthy
通讯作者：
D. Spink;J. Johnson;S. Connor;K. Aldous;J. Gierthy
野生动物非传染性疾病
影响因子：
3
作者：
A. Fairbrother;L. N. Locke;G. Hoff
通讯作者：
G. Hoff
影响因子：
13.5
作者：
S. Safe
通讯作者：
S. Safe
电子屏幕测定作为识别雌激素的工具：雌激素环境污染物的更新。
影响因子：
10.4
作者：
通讯作者：
--
影响因子：
4.5
作者：
Anderson, MJ;Miller, MR;Hinton, DE
通讯作者：
Hinton, DE