Instituto de Toxicologia e Farmacologia, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, CEP 90619-900, Brazil; Programa de Pós-Graduação em Medicina: Ciências Médicas, Universidade Federal do Rio Grande do Sul, Porto Alegre, CEP 90035-003, Brazil.
Toxicol Appl Pharmacol. 2013 Nov 1;272(3):681-9. doi: 10.1016/j.taap.2013.08.001. Epub 2013 Aug 8.
The use of zebrafish (Danio rerio) is increasing as an intermediate preclinical model, to prioritize drug candidates for mammalian testing. As the immune system of the zebrafish is quite similar to that of mammals, models of inflammation are being developed for the screening of new drugs. The characterization of these models is crucial for studies that seek for mechanisms of action and specific pharmacological targets. It is well known that copper is a metal that induces damage and cell migration to hair cells of lateral line of zebrafish. Extracellular nucleotides/nucleosides, as ATP and adenosine (ADO), act as endogenous signaling molecules during tissue damage by exerting effects on inflammatory and immune responses. The present study aimed to characterize the inflammatory status, and to investigate the involvement of the purinergic system in copper-induced inflammation in zebrafish larvae. Fishes of 7 days post-fertilization were exposed to 10 μM of copper for a period of 24 h. The grade of oxidative stress, inflammatory status, copper uptake, the activity and the gene expression of the enzymes responsible for controlling the levels of nucleotides and adenosine were evaluated. Due to the copper accumulation in zebrafish larvae tissues, the damage and oxidative stress were exacerbated over time, resulting in an inflammatory process involving IL-1β, TNF-α, COX-2 and PGE2. Within the purinergic system, the mechanisms that control the ADO levels were the most involved, mainly the reactions performed by the isoenzyme ADA 2. In conclusion, our data shed new lights on the mechanisms related to copper-induced inflammation in zebrafish larvae.
斑马鱼（Danio rerio）作为一种中间临床前模型，其应用正在增加，以优先选择候选药物进行哺乳动物测试。由于斑马鱼的免疫系统与哺乳动物非常相似，因此正在开发炎症模型来筛选新药。这些模型的特征对于寻求作用机制和特定药理学靶点的研究至关重要。众所周知，铜是一种会导致斑马鱼侧线毛细胞损伤和迁移的金属。细胞外核苷酸/核苷，如 ATP 和腺苷（ADO），在组织损伤时作为内源性信号分子发挥作用，影响炎症和免疫反应。本研究旨在表征炎症状态，并研究嘌呤能系统在铜诱导的斑马鱼幼虫炎症中的参与。受精后 7 天的鱼类暴露于 10 μM 的铜 24 小时。评估氧化应激程度、炎症状态、铜摄取、控制核苷酸和腺苷水平的酶的活性和基因表达。由于铜在斑马鱼幼虫组织中的积累，随着时间的推移，损伤和氧化应激加剧，导致涉及 IL-1β、TNF-α、COX-2 和 PGE2 的炎症过程。在嘌呤能系统中，控制 ADO 水平的机制最相关，主要是由同工酶 ADA 2 执行的反应。总之，我们的数据为斑马鱼幼虫铜诱导炎症相关的机制提供了新的认识。
