Alves Rodrigo F, Lopes Célia, Rocha Eduardo, Madureira Tânia Vieira
Team of Animal Morphology and Toxicology, Interdisciplinary Centre of Marine and Environmental Research (CIIMAR/CIMAR), University of Porto (U.Porto), Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos s/n, 4450-208 Matosinhos, Portugal.
Laboratory of Histology and Embryology, Department of Microscopy, ICBAS-School of Medicine and Biomedical Sciences, University of Porto (U.Porto), Rua Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
J Xenobiot. 2024 Aug 6;14(3):1064-1078. doi: 10.3390/jox14030060.
Three-dimensional (3D) fish hepatocyte cultures are promising alternative models for replicating in vivo data. Few studies have attempted to characterise the structure and function of fish 3D liver models and illustrate their applicability. This study aimed to further characterise a previously established spheroid model obtained from juvenile brown trout () primary hepatocytes under estrogenic stimulation. The spheroids were exposed for six days to environmentally relevant concentrations of 17α-ethinylestradiol-EE2 (1-100 ng/L). The mRNA levels of peroxisome ( and ), lipid metabolism (, , and ), and estrogen-related (, , , , and ) target genes were evaluated by quantitative real-time polymerase chain reaction. Immunohistochemistry was used to assess Vtg and ZP protein expressions. At the highest EE2 concentration, and genes were significantly upregulated. The remaining target genes were not significantly altered by EE2. Vtg and ZP immunostaining was consistently increased in spheroids exposed to 50 and 100 ng/L of EE2, whereas lower EE2 levels resulted in a weaker signal. EE2 did not induce significant changes in the spheroids' viability and morphological parameters. This study identified EE2 effects at environmentally relevant doses in trout liver spheroids, indicating its usefulness as a proxy for in vivo impacts of xenoestrogens.
三维（3D）鱼类肝细胞培养是复制体内数据的有前景的替代模型。很少有研究试图表征鱼类3D肝脏模型的结构和功能并说明其适用性。本研究旨在进一步表征先前建立的在雌激素刺激下从幼年褐鳟（）原代肝细胞获得的球体模型。将球体暴露于环境相关浓度的17α-乙炔雌二醇-EE2（1-100 ng/L）中六天。通过定量实时聚合酶链反应评估过氧化物酶体（和）、脂质代谢（、和）以及雌激素相关（、、、和）靶基因的mRNA水平。免疫组织化学用于评估Vtg和ZP蛋白表达。在最高EE2浓度下，和基因显著上调。其余靶基因未被EE2显著改变。在暴露于50和100 ng/L EE2的球体中，Vtg和ZP免疫染色持续增加，而较低的EE2水平导致信号较弱。EE2未诱导球体活力和形态参数的显著变化。本研究确定了环境相关剂量的EE2对鳟鱼肝脏球体的影响，表明其作为异源雌激素体内影响替代物的有用性。
