近年来，不同药物对甲状腺系统的潜在毒性引起了越来越多的关注。本研究旨在评估帕唑帕尼和阿昔替尼这两种广泛应用于临床的抗肿瘤药物在斑马鱼模型中对甲状腺功能的毒性作用。我们使用商业酶联免疫吸附试验（ELISA）套装测量了甲状腺相关激素的水平。采用全蛋白原位杂交（WISH）分析技术检测目标基因表达变化。使用共聚焦显微镜评估了转基因Tg（tg: EGFP）鱼株中甲状腺的形态。通过定量实时聚合酶链反应（RT-qPCR）验证了关键基因的相对mRNA表达水平。使用血红素-伊红染色方法在光学显微镜下定量评估了滋养层大小和数量。结果显示，在帕唑帕尼或阿昔替尼中孵化96小时的斑马鱼胚胎中，发育和存活受到明显影响，并伴随着甲状腺内分泌系统的显著紊乱（如甲状腺刺激素（TSH）含量增加及三碘甲状腺原氨酸（T3）和甲状腺素（T4）含量降低，以及与下丘脑-垂体-甲状腺（HPT）轴相关的基因转录变化）。此外，根据帕唑帕尼和阿昔替尼处理的斑马鱼胚胎的全蛋白原位杂交染色和组织病理学检查，我们观察到Tg（tg: EGFP）斑马鱼转基因株中甲状腺滋养层发育明显异常。总的来说，这些发现表明，帕唑帕尼和阿昔替尼可能通过影响HPT轴的调节，部分地对甲状腺的发育和功能产生毒性效应。因此，我们认为在它们的临床应用中应更加重视帕唑帕尼和阿昔替尼的潜在甲状腺毒性作用。版权所有 ©2023 Yang, Tu, Zhang, Xie, Dong, Jing, Chen, Wei and Song.

In recent years, the potential toxicities of different pharmaceuticals toward the thyroid system have received increasing attention. In this study, we aim to evaluate the toxic effects of pazopanib and axitinib, two anti-tumor drugs with widespread clinical use, on thyroid function in the zebrafish model.We measured levels of thyroid-related hormones using the commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit. Whole-mount in situ hybridization (WISH) analysis was employed to detect target gene expression changes. Morphology of the thyroid were evaluated by using transgenic Tg (tg: EGFP) fish line under a confocal microscope. The relative mRNA expression of key genes was verified through quantitative real-time polymerase chain reaction (RT‒qPCR). The size and number of the follicles was quantified whereby Hematoxylin-Eosin (H & E) staining under a light microscope.The results revealed that fertilized zebrafish embryos were incubated in pazopanib or axitinib for 96 hours, development and survival were significantly affected, which was accompanied by significant disturbances in thyroid endocrine system (e.g., increased thyroid-stimulating hormone (TSH) content and decreased triiodothyronine (T3) and thyroxine (T4) content, as well as transcription changes of genes associated with the hypothalamus-pituitary-thyroid (HPT) axis. Moreover, based on whole-mount in situ hybridization staining of tg and histopathological examination of zebrafish embryos treated with pazopanib and axitinib, we observed a significantly abnormal development of thyroid follicles in the Tg (tg: EGFP) zebrafish transgenic line.Collectively, these findings indicate that pazopanib and axitinib may have toxic effects on thyroid development and function, at least partially, by influencing the regulation of the HPT axis. Thus, we believe that the potential thyroid toxicities of pazopanib and axitinib in their clinical applications should receive greater attention.Copyright © 2023 Yang, Tu, Zhang, Xie, Dong, Jing, Chen, Wei and Song.