Guo Shengya, Zhang Xiaoxi, Zhang Yong, Chen Xinghui, Zhang Yiwen, Cao Bingbing, Xia Daozong
College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China; Hunter Biotechnology, Inc., Hangzhou, 310051, China.
College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China.
Chemosphere. 2023 Oct 27:140561. doi: 10.1016/j.chemosphere.2023.140561.
Lead (Pb) contamination is a worldwide public health threaten. Besides close restraint of lead exposure, it's emergency to discover compounds that could help to cue toxicities caused by lead. Zebrafish embryos and early larvae can serve as valuable screening tools in early pre-clinical phase of drug screening and research. In order to establish a zebrafish lead poisoning model that could be used for drug screening and research, zebrafish embryos at 6 h post-fertilization (hpf) were treated with lead at different concentrations by soaking intermittently, raised in lead work solution at night while in fish water during the day. After treated for 90 h, death and severe trunk curvature were observed on zebrafish in 640 μM group, obvious dysplasia, blood toxicity, excessive reactive oxygen species (ROS), severe neurotoxicity, such as shorter length of peripheral motor neurons, neuronal apoptosis, and axonal injury, and neurobehavior impairment were induced by lead at 80, 160 and 320 μM, similar to phenotypes reported in rodent. Moreover, the mRNA level of genes related to neurodevelopment, memory, and antioxidation were significantly down regulated, and apoptosis-related genes were up regulated, consistent to zebrafish phenotypic change. Finally, zebrafish were intermittently exposed to 80 μM lead solution to establish the lead poisoning model, and the efficacy of a safe chelating agent Meso-2,3-Dimercaptosuccinic acid (DMSA) was tested at a series of concentrations to validate the zebrafish model. The result showed concentration-dependent decrease of lead content in zebrafish in DMSA treated groups compared with model group. The above data fully demonstrated a zebrafish model of lead poisoning suitable for drug screening was successfully developed, which was expected to provide a rapid and economic tools for discovering antidotes of lead and drugs of neuroprotection.
铅（Pb）污染是一个全球性的公共卫生威胁。除了严格限制铅暴露外，发现有助于提示铅所致毒性的化合物也很紧急。斑马鱼胚胎和早期幼体可作为药物筛选和研究临床前早期阶段有价值的筛选工具。为了建立可用于药物筛选和研究的斑马鱼铅中毒模型，对受精后6小时（hpf）的斑马鱼胚胎进行不同浓度的铅间歇浸泡处理，夜间饲养于含铅工作溶液中，白天饲养于鱼水中。处理90小时后，640μM组斑马鱼出现死亡和严重的躯干弯曲，80、160和320μM的铅诱导了明显的发育异常、血液毒性、过量活性氧（ROS）、严重的神经毒性，如外周运动神经元长度缩短、神经元凋亡和轴突损伤以及神经行为损害，类似于啮齿动物中报道的表型。此外，与神经发育、记忆和抗氧化相关的基因的mRNA水平显著下调，而凋亡相关基因上调，与斑马鱼的表型变化一致。最后，将斑马鱼间歇暴露于80μM铅溶液中建立铅中毒模型，并测试一系列浓度的安全螯合剂中-2,3-二巯基丁二酸（DMSA）的疗效以验证斑马鱼模型。结果显示，与模型组相比，DMSA处理组斑马鱼体内铅含量呈浓度依赖性降低。上述数据充分证明成功建立了适合药物筛选的斑马鱼铅中毒模型，有望为发现铅的解毒剂和神经保护药物提供一种快速且经济的工具。
