Department of Anesthesiology, University of Illinois, Chicago, Illinois 60612.
Research and Development, Jesse Brown VA Medical Center, Chicago, Illinois 60612.
Toxicol Sci. 2017 Sep 1;159(1):224-237. doi: 10.1093/toxsci/kfx134.
Superwarfarins are very long-lasting rodenticides effective in warfarin-resistant rodents at extremely low doses. The consequences of chronic superwarfarin levels in tissues, due to biological half-lives on the order of 20 days, have not been examined. We now characterized the neurological effects of brodifacoum (BDF), one of the most widely used superwarfarins, in adult male Sprague Dawley rats. Dosing curves established the acute oral lethal dose for BDF as 221 ± 14 μg/kg. Measurement of tissue BDF levels showed accumulation throughout the body, including the central nervous system, with levels diminishing over several days. Immunocytochemical staining showed that both astrocyte and microglial activation was increased 4 days after BDF administration, as were levels of carbonylated proteins, and neuronal damage assessed by fluorojade B staining. Direct toxic effects of BDF on neurons and glia were observed using enriched cultures of cerebellar neurons and cortical astrocytes. Proteomic analysis of cerebellar lysates revealed that BDF altered expression of 667 proteins in adult rats. Gene ontology and pathway analysis identified changes in several functional pathways including cell metabolism, mitochondria function, and RNA handling with ribosomal proteins comprising the largest group. In vitro studies using primary astrocytes showed that BDF suppressed de novo protein synthesis. These findings demonstrate that superwarfarin accumulation increases indices of neuroinflammation and neuropathology in adult rodents, suggesting that methods which minimize BDF toxicity may not address delayed neurological sequelae.
超级杀鼠剂是非常长效的杀鼠剂,对具有抗华法林特性的啮齿类动物只需极低剂量即可生效。由于其生物半衰期约为 20 天,组织中慢性超级杀鼠剂水平的后果尚未得到检验。我们现在研究了其中一种广泛使用的超级杀鼠剂——溴敌隆(BDF)对成年雄性 Sprague Dawley 大鼠的神经毒性作用。剂量曲线确定了 BDF 的急性口服致死剂量为 221±14μg/kg。组织 BDF 水平的测量表明,BDF 在全身(包括中枢神经系统)积累,数天后水平逐渐降低。免疫细胞化学染色显示,BDF 给药 4 天后,星形胶质细胞和小胶质细胞的激活均增加,羰基化蛋白水平以及用氟罗丹明 B 染色评估的神经元损伤也增加。使用小脑神经元和皮质星形胶质细胞的富集培养物观察到 BDF 对神经元和神经胶质的直接毒性作用。小脑裂解物的蛋白质组学分析显示,BDF 改变了成年大鼠中 667 种蛋白质的表达。基因本体论和途径分析确定了几个功能途径的变化,包括细胞代谢、线粒体功能和 RNA 处理,其中核糖体蛋白构成最大的一组。使用原代星形胶质细胞的体外研究表明,BDF 抑制了新蛋白质的合成。这些发现表明,超级杀鼠剂的积累增加了成年啮齿动物的神经炎症和神经病理学指标,这表明可能无法通过最小化 BDF 毒性的方法来解决延迟的神经后遗症。