BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany.
Regul Toxicol Pharmacol. 2012 Jul;63(2):259-78. doi: 10.1016/j.yrtph.2012.04.004. Epub 2012 Apr 24.
Most endocrine disruptors interact with hormone receptors or steroid biosynthesis and metabolism, thereby modifying the physiological function of endogenous hormones. Here, we present an alternative testing paradigm for detection of endocrine modes of action that replace and reduce animal testing through refinement. Receptor mediated endocrine effects were assessed using the yeast-based receptor-mediated transcriptional activation YES/YAS assays and effects on steroid hormone biosynthesis were assessed using the human cell line H295R in the steroidogenesis assay. In our testing paradigm we propose to complement the in vitro assays with a single in vivo repeated dose study in which plasma samples are analyzed for their metabolome profile in addition to classical parameters such as histopathology. The combination of these methods does not only contribute to refinement and reduction of animal testing, but also has significantly increased the efficient allocation of resources and allows for a sound assessment of the endocrine disruption potential of compounds. Thus, this proposal constitutes a potentially attractive alternative to EPA's Endocrine Disruptor Screening Program to identify mammalian, systemic endocrine modes of action. Data on 14 reference substances for which the in vitro YES/YAS and steroidogenesis assays and the in vivo metabolome analysis were performed to assess their putative endocrine modes of action are presented here.
大多数内分泌干扰物通过与激素受体或类固醇生物合成和代谢相互作用，从而改变内源性激素的生理功能。在这里，我们提出了一种替代的测试范例，用于检测内分泌作用模式，通过精细化替代和减少动物测试。使用基于酵母的受体介导转录激活 YES/YAS 测定来评估受体介导的内分泌效应，并且使用类固醇生成测定中的人细胞系 H295R 评估对类固醇激素生物合成的影响。在我们的测试范例中，我们建议将体外测定与单次重复剂量的体内研究相结合，除了经典参数如组织病理学外，还分析血浆样本的代谢组特征。这些方法的组合不仅有助于细化和减少动物测试，而且还显著增加了资源的有效分配，并允许对化合物的内分泌干扰潜力进行合理评估。因此，本建议构成了 EPA 的内分泌干扰物筛选计划的一种有吸引力的替代方案，用于识别哺乳动物、系统性的内分泌作用模式。本文介绍了对 14 种参考物质进行的体外 YES/YAS 和类固醇生成测定以及体内代谢组分析的数据，以评估它们潜在的内分泌作用模式。
