Laboratory of Cell Biology and Histology, Department of Zoology and Animal Cell Biology, Faculty of Science and Technology, University of the Basque Country (UPV/EHU), Sarriena z/g, E-48940 Leioa, Basque Country, Spain; Research Centre for Experimental Marine Biology and Biotechnology PIE, University of the Basque Country (UPV/EHU), Areatza z/g, E-48620 Plentzia, Basque Country, Spain.
Sci Total Environ. 2014 Jan 1;466-467:1-10. doi: 10.1016/j.scitotenv.2013.06.060. Epub 2013 Jul 25.
Commercial OP and NP are complex isomer mixtures that can be individually present in the environment, showing different estrogenic potencies. The aims of this study were to establish the estrogenic potency of some AP isomers in comparison to the commercial NP (cNP) mixture in vitro and to investigate in vivo their possible effects during the embryo and larval development of zebrafish. An in vitro estrogen receptor-based recombinant yeast assay was used to test the estrogenicity of specific AP isomers (22-OP, 33-OP, 22-NP, 33-NP and 363-NP) and cNP. The EC₅₀ was in the range of 0.6-7.7 mg/L. Both OP isomers and 363-NP exhibited higher estrogenic activity than cNP. For in vivo experiments, one-day postfertilisation (dpf) embryos were exposed to cNP (50, 250 and 500 μg/L), 363-NP and 33-OP (50 μg/L), 17β-estradiol (100 ng/L) and DMSO (0.01% v/v) for 4weeks. After exposure fish were maintained for 2 weeks in clean water in order to evaluate a possible recovery. Fish of groups exposed to cNP and 363-NP were the last to hatch. Histological alterations were not observed after 7, 28 or 42 dpf. Exposure to 33-OP increased transcriptional levels of erα, vtg and cyp19a1b genes. However, transcriptional response in E2 exposure was observed at later stages and with higher fold induction levels. Exposure to cNP decreased levels of erα whereas increased levels of rxrγ and cyp19a1b. Exposure to 363-NP did not cause changes in transcriptional levels of studied genes. The differences in response of the OP isomer compared to the NP isomer in zebrafish could be related to the rapid decay in concentration of the latter.
商业性邻苯二甲酸二辛酯（OP）和壬基酚（NP）是复杂的同分异构体混合物，它们可单独存在于环境中，表现出不同的雌激素活性。本研究的目的是在体外建立某些邻苯二甲酸酯（AP）异构体相对于商业性 NP（cNP）混合物的雌激素活性，并研究它们在斑马鱼胚胎和幼虫发育过程中的潜在影响。使用基于雌激素受体的重组酵母测定法来测试特定 AP 异构体（22-OP、33-OP、22-NP、33-NP 和 363-NP）和 cNP 的雌激素活性。EC₅₀ 值在 0.6-7.7 mg/L 范围内。OP 异构体和 363-NP 均表现出比 cNP 更高的雌激素活性。对于体内实验，将 1 天龄受精卵（dpf）暴露于 cNP（50、250 和 500 μg/L）、363-NP 和 33-OP（50 μg/L）、17β-雌二醇（100 ng/L）和 DMSO（0.01%v/v）中 4 周。暴露后，鱼在清洁水中维持 2 周，以评估可能的恢复情况。暴露于 cNP 和 363-NP 的鱼最后孵化。在 7、28 或 42 dpf 时未观察到组织学改变。暴露于 33-OP 增加了 erα、vtg 和 cyp19a1b 基因的转录水平。然而，在后期阶段和更高的倍数诱导水平观察到 E2 暴露的转录反应。暴露于 cNP 降低了 erα 的水平，而增加了 rxrγ 和 cyp19a1b 的水平。暴露于 363-NP 不会导致研究基因的转录水平发生变化。与 NP 异构体相比，OP 异构体在斑马鱼中的反应差异可能与其浓度的快速衰减有关。
