Magalhães Francisco Ernani Alves, de Sousa Caio Átila Prata Bezerra, Santos Sacha Aubrey Alves Rodrigues, Menezes Renata Barbosa, Batista Francisco Lucas Alves, Abreu Ângela Oliveira, de Oliveira Messias Vital, Moura Luiz Francisco Wemmenson Gonçalves, Raposo Ramon da Silva, Campos Adriana Rolim
1 LBPNB (Laboratório de Bioprospecção de Produtos Naturais e Biotecnologia), Universidade Estadual do Ceará (UECE) , Tauá, Ceará, Brazil .
2 NUBEX (Núcleo de Biologia Experimental), Universidade de Fortaleza (UNIFOR) , Fortaleza, Ceará, Brazil .
Zebrafish. 2017 Oct;14(5):422-429. doi: 10.1089/zeb.2017.1436. Epub 2017 Jul 13.
The zebrafish (Danio rerio) has been proposed as a low-cost and simple alternative to the use of higher vertebrates in laboratory research on novel compounds with antinociceptive potential. In this study, we tested adult zebrafish (Danio rerio) as an alternative behavioral model of formalin-induced nociception. We evaluated the nociceptive effect of 0.1% formalin (3 or 5 μL; intramuscularly [i.m.]), applied into the tail or lips, on locomotor activity, using as parameter the number of times the fish crossed the lines between the quadrants of a glass Petri dish during the neurogenic stage (0-5 min) and the inflammatory stage (15-30 min). The behavioral model was validated by testing the antinociceptive effect of morphine and indomethacin (standard analgesic drugs used in the formalin test of rodents). We also tested whether the effect of morphine could be modulated by naloxone, an opioid antagonist. The effect of morphine and indomethacin on zebrafish locomotor behavior was evaluated with the open field test. The white/black test was used to rule out the anxiolytic effect of 0.1% formalin injected into the tail on adult zebrafish. Formalin (0.1%; 3 and 5 μL injected into the tail) increased significantly the nociceptive behavior of the adult zebrafish in both stages (p < 0.001 vs. control). Morphine and indomethacin (both 0.2 mg/mL; 20 μL; intraperitoneally [i.p.]) significantly inhibited nociception induced with formalin (5 μL injected i.m. into the tail) in both stages (p < 0.001). Naloxone blocked the antinociceptive effect of morphine. No influence on locomotion was observed. Locally administered formalin (injected into the tail) induced nociception, but not anxiety. The results suggest that the adult zebrafish behavioral model is a feasible alternative to more conventional laboratory models used in research on novel compounds with antinociceptive potential.
斑马鱼（Danio rerio）已被提议作为一种低成本且简便的替代方法，用于在实验室中对具有抗伤害感受潜力的新型化合物进行研究时替代高等脊椎动物。在本研究中，我们测试了成年斑马鱼（Danio rerio）作为福尔马林诱导伤害感受的替代行为模型。我们评估了将0.1%福尔马林（3或5μL；肌肉注射[i.m.]）注入尾巴或嘴唇对运动活动的伤害感受作用，以在神经源性阶段（0 - 5分钟）和炎症阶段（15 - 30分钟）期间鱼穿过玻璃培养皿象限之间线条的次数作为参数。通过测试吗啡和吲哚美辛（啮齿动物福尔马林试验中使用的标准镇痛药）的抗伤害感受作用来验证该行为模型。我们还测试了吗啡的作用是否可被阿片类拮抗剂纳洛酮调节。通过旷场试验评估吗啡和吲哚美辛对斑马鱼运动行为的影响。使用白/黑试验来排除将0.1%福尔马林注入尾巴对成年斑马鱼的抗焦虑作用。福尔马林（0.1%；3和5μL注入尾巴）在两个阶段均显著增加了成年斑马鱼的伤害感受行为（与对照组相比，p < 0.001）。吗啡和吲哚美辛（均为0.2mg/mL；20μL；腹腔注射[i.p.]）在两个阶段均显著抑制了福尔马林（5μL肌肉注射入尾巴）诱导的伤害感受（p < 0.001）。纳洛酮阻断了吗啡的抗伤害感受作用。未观察到对运动的影响。局部注射福尔马林（注入尾巴）诱导了伤害感受，但未诱导焦虑。结果表明，成年斑马鱼行为模型是研究具有抗伤害感受潜力的新型化合物时更传统实验室模型的可行替代方法。
