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首页> 外文期刊>Environmental toxicology and chemistry >UTILITY OF IN VITRO TEST METHODS TO ASSESS THE ACTIVITY OF XENOESTROGENS IN FISH
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UTILITY OF IN VITRO TEST METHODS TO ASSESS THE ACTIVITY OF XENOESTROGENS IN FISH

机译：体外测试方法评估鱼类异种雌激素活性的实用性

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摘要

The results of the present study have demonstrated the utility of an estrogen receptor (ER) competitive ligand-binding assay, a hepatocyte vitellogenin (VTG) induction bioassay, and an ER reporter gene bioassay in characterizing the activity of model estrogen agonists (17β-estradiol [E_2], ethynylestradiol, and nonylphenol) and antagonists (tamoxifen and ZM 189154) in rainbow trout (Oncorhynchus mykiss). The in vitro results were validated against in vivo trout waterborne exposures to E_2 and tamoxifen. The results showed that all three in vitro assays were capable of detecting the hormonal activities of the selected model compounds in a dose-dependent manner, with the exception of nonylphenol in the ER reporter gene bioassay. However, the relative potency rankings of the model compounds were not consistent between these assays, which suggests that the relative potencies obtained within assays may have limited predictive value between assays. Discrepancies in potencies most likely can be attributed to the different levels of cellular organization in each assay. In addition to model compounds, we also evaluated the responses of the ER-binding assay and the hepatocyte VTG induction bioassay to complex mixtures associated with endocrine effects in fish, specifically extracts of pulp mill effluent. Of the 14 pulp mill effluent extracts tested, only six showed activity in both assays, whereas the remaining eight samples showed activity in only one of the two assays. The hepatocyte VTG induction bioassay consistently showed that the pulp mill effluent extracts were antiestrogenic, which to our knowledge has not been reported in previous studies. Collectively, these results suggest that a combination of in vitro assays that depend on differing endpoints is required to identify potential xenoestrogens and to characterize their modes of action.
机译：本研究的结果证明了雌激素受体（ER）竞争性配体结合测定，肝细胞卵黄蛋白原（VTG）诱导生物测定和ER报告基因生物测定在表征模型雌激素激动剂（17β-雌二醇）活性方面的实用性虹鳟鱼（Oncorhynchus mykiss）中的[E_2]，乙炔雌二醇和壬基酚）和拮抗剂（他莫昔芬和ZM 189154）。体外结果对鳟鱼水体暴露于E_2和他莫昔芬的效果进行了验证。结果表明，除了ER报告基因生物测定中的壬基酚外，所有三种体外测定均能够以剂量依赖的方式检测所选模型化合物的激素活性。但是，在这些测定之间，模型化合物的相对效价等级不一致，这表明在测定中获得的相对效价可能在测定之间具有有限的预测价值。效能差异很可能归因于每种测定中细胞组织水平的不同。除模型化合物外，我们还评估了ER结合测定和肝细胞VTG诱导生物测定对与鱼类内分泌作用相关的复杂混合物（特别是制浆厂废水的提取物）的响应。在测试的14种制浆厂废液提取物中，只有两种在两种测定中均显示出活性，而其余八种样品仅在两种测定中之一中显示出活性。肝细胞VTG诱导生物测定始终表明，制浆厂的废液提取物具有抗雌激素作用，据我们所知，以前的研究中尚未对此进行报道。总的来说，这些结果表明，需要使用依赖于不同终点的体外测定方法的组合来鉴定潜在的异雌激素并表征其作用方式。

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