Beijing Key Laboratory of Fishery Biotechnology, Beijing Fisheries Research Institute, Beijing 100068, China.
Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Int J Mol Sci. 2018 Nov 8;19(11):3516. doi: 10.3390/ijms19113516.
Notwithstanding the widespread use and promising clinical value of chemotherapy, the pharmacokinetics, toxicology, and mechanism of mitoxantrone remains unclear. To promote the clinical value in the treatment of human diseases and the exploration of potential subtle effects of mitoxantrone, zebrafish embryos were employed to evaluate toxicity with validated reference genes based on independent stability evaluation programs. The most stable and recommended reference gene was , followed by , for the 48 h post fertilization (hpf) zebrafish embryo mitoxantrone test, while both and were recommended as reference genes for the 96 hpf zebrafish embryo mitoxantrone test. With as an internal control, we analyzed the mRNA levels of representative hepatotoxicity biomarkers, including , , , , cardiotoxicity biomarker , and neurotoxicity biomarker in the 48 hpf embryo mitoxantrone test. The mRNA levels of , , and increased significantly in 10 and 50 μg/L mitoxantrone-treated 48 hpf embryos, while the transcript levels of decreased in a dose-dependent manner, indicating that mitoxantrone induced hepatotoxicity and neurotoxicity. Liver hematoxylin⁻eosin staining and the spontaneous movement of embryos confirmed the results. Thus, the present research suggests that mitoxantrone induces toxicity during the development of the liver and nervous system in zebrafish embryos and that is recommended as a potential biomarker for hepatotoxicity in zebrafish embryos. Additionally, is proposed as a reference gene for the 48 hpf zebrafish embryo mitoxantrone toxicity test, while and are proposed as that for the 96 hpf test.
尽管化疗的应用广泛且具有有前景的临床价值，但米托蒽醌的药代动力学、毒理学和作用机制仍不清楚。为了促进米托蒽醌在治疗人类疾病中的临床价值，并探索其潜在的微妙作用，本研究采用斑马鱼胚胎，基于独立稳定性评估程序，使用经过验证的参考基因来评估毒性。对于 48 小时受精后（hpf）的斑马鱼胚胎米托蒽醌试验，最稳定和推荐的参考基因是 ，其次是 ，而对于 96 hpf 的斑马鱼胚胎米托蒽醌试验，推荐使用 和 作为参考基因。以 为内参，我们分析了代表肝毒性生物标志物的 mRNA 水平，包括 、 、 、 、心脏毒性生物标志物 和神经毒性生物标志物 在 48 hpf 胚胎米托蒽醌试验中的表达。在 10 和 50 μg/L 米托蒽醌处理的 48 hpf 胚胎中， 、 和 的 mRNA 水平显著增加，而 的转录水平呈剂量依赖性下降，表明米托蒽醌诱导了肝毒性和神经毒性。肝苏木精-伊红染色和胚胎自发运动证实了这一结果。因此，本研究表明米托蒽醌在斑马鱼胚胎肝脏和神经系统发育过程中引起毒性， 被推荐为斑马鱼胚胎肝毒性的潜在生物标志物。此外， 被推荐为 48 hpf 斑马鱼胚胎米托蒽醌毒性试验的参考基因， 或 被推荐为 96 hpf 试验的参考基因。
