Institut National de l'Environnement Industriel et des Risques (INERIS), Unité Écotoxicologie in vitro et in vivo, Verneuil-en-Halatte, France; INRA, UMR1331, Toxalim, Research Centre in Food Toxicology, Toulouse, France; Université de Toulouse, INP, UMR 1331 TOXALIM, Toulouse, France.
Institut National de l'Environnement Industriel et des Risques (INERIS), Unité Écotoxicologie in vitro et in vivo, Verneuil-en-Halatte, France.
Ecotoxicol Environ Saf. 2017 Aug;142:150-156. doi: 10.1016/j.ecoenv.2017.04.009. Epub 2017 Apr 11.
Bisphenol A (BPA) is a widely used chemical that has been extensively studied as an endocrine-disrupting chemical (EDC). Other bisphenols sharing close structural features with BPA, are increasingly being used as alternatives, increasing the need to assess associated hazards to the endocrine system. In the present study, the estrogenic activity of BPA, bisphenol S (BPS) and bisphenol F (BPF) was assessed by using a combination of zebrafish-specific mechanism-based in vitro and in vivo assays. The three bisphenols were found to efficiently transactivate all zebrafish estrogen receptor (zfER) subtypes in zebrafish hepatic reporter cell lines (ZELH-zfERs). BPA was selective for zfERα while BPS and BPF were slightly more potent on zfERβ subtypes. We further documented the estrogenic effect in vivo by quantifying the expression of brain aromatase using a transgenic cyp19a1b-GFP zebrafish embryo assay. All three bisphenols induced GFP in a concentration-dependent manner. BPS only partially induced brain aromatase at the highest tested concentrations (>30µM) while BPA and BPF strongly induced GFP, in an ER-dependent manner, at 1-10µM. Furthermore, we show that BPF strongly induced vitellogenin synthesis in adult male zebrafish. Overall, this study demonstrates the estrogenic activity of BPA, BPF and BPS in different cell- and tissue-contexts and at different stages of development. Differences between in vitro and in vivo responses are discussed in light of selective ER activation and the fate of the compounds in the models. This study confirms the relevance of combining cellular and whole-organism bioassays in a unique model species for the hazard assessment of candidate EDCs.
双酚 A（BPA）是一种广泛使用的化学物质，作为一种内分泌干扰化学物质（EDC），已被广泛研究。其他与 BPA 具有密切结构特征的双酚类物质，作为替代品的使用越来越多，这增加了评估其对内分泌系统相关危害的必要性。在本研究中，采用结合斑马鱼特异性基于机制的体外和体内测定法，评估了 BPA、双酚 S（BPS）和双酚 F（BPF）的雌激素活性。发现这三种双酚类物质能有效地在斑马鱼肝报告细胞系（ZELH-zfERs）中转激活所有斑马鱼雌激素受体（zfER）亚型。BPA 对 zfERα 具有选择性，而 BPS 和 BPF 对 zfERβ 亚型的活性略强。我们进一步通过使用 Cyp19a1b-GFP 转基因斑马鱼胚胎测定法量化脑芳香酶的表达，在体内记录了雌激素效应。所有三种双酚类物质均以浓度依赖的方式诱导 GFP。BPS 仅在最高测试浓度（>30µM）下部分诱导脑芳香酶，而 BPA 和 BPF 则以 ER 依赖性方式在 1-10µM 时强烈诱导 GFP。此外，我们还表明 BPF 强烈诱导成年雄性斑马鱼合成卵黄蛋白原。总体而言，这项研究证明了 BPA、BPF 和 BPS 在不同细胞和组织背景以及在不同发育阶段的雌激素活性。结合细胞和整体生物测定法，在独特的候选 EDC 危险评估模式物种中，讨论了体外和体内反应之间的差异。
