Department of Comparative Biomedical Sciences, Center for Environmental Health Sciences, Mississippi State University, College of Veterinary Medicine, Mississippi State, United States, USA.
J Biochem Mol Toxicol. 2021 Jun;35(6):1-10. doi: 10.1002/jbt.22755. Epub 2021 Mar 8.
Past assassinations and terrorist attacks demonstrate the need for a more effective antidote against nerve agents and other organophosphates (OP) that cause brain damage through inhibition of acetylcholinesterase (AChE). Our lab has invented a platform of phenoxyalkyl pyridinium oximes (US patent 9,277,937) that demonstrate the ability to cross the blood-brain barrier in in vivo rat tests with a sarin surrogate nitrophenyl isopropyl methylphosphonate (NIMP) and provide evidence of brain penetration by reducing cessation time of seizure-like behaviors, accumulation of glial fibrillary acidic protein (GFAP), and hippocampal neuropathology, as opposed to the currently approved oxime, 2-pyridine aldoxime methyl chloride (2-PAM). Using two of the novel oximes (Oximes 1 and 20), this project examined whether gene expression changes might help explain this protection. Expression changes in the piriform cortex were examined using polymerase chain reaction arrays for inflammatory cytokines and receptors. The hippocampus was examined via quantitative polymerase chain reaction for the expression of immediate-early genes involved in brain repair (Bdnf), increasing neurotoxicity (Fos), and apoptosis control (Jdp2, Bcl2l1, Bcl2l11). In the piriform cortex, NIMP significantly stimulated expression for the macrophage inflammatory proteins CCL4, IL-1A, and IL-1B. Oxime 20 by itself elicited the most changes. When it was given therapeutically post-NIMP, the largest change occurred: a 310-fold repression of the inflammatory cytokine, CCL12. In the hippocampus, NIMP increased the expression of the neurotoxicity marker Fos and decreased the expression of neuroprotective Bdnf and antiapoptotic Bcl2l1. Compared with 2-PAM, Oxime 20 stimulated Bcl2l1 expression more and returned expression closer to the vehicle control values.
过去的暗杀和恐怖袭击事件表明,需要一种更有效的解毒剂来对抗神经毒剂和其他有机磷化合物(OP),这些化合物通过抑制乙酰胆碱酯酶(AChE)而导致脑损伤。我们的实验室发明了一种苯氧乙基吡啶𬭩肟(美国专利 9,277,937)平台,该平台在体内大鼠试验中用沙林替代物硝苯基异丙基甲基膦酸酯(NIMP)证明了穿过血脑屏障的能力,并通过减少癫痫样行为的停止时间、神经胶质纤维酸性蛋白(GFAP)的积累和海马神经病理学,提供了脑穿透的证据,而不是目前批准的肟,2-吡啶醛肟甲基氯化物(2-PAM)。使用两种新型肟(Oxime 1 和 20),本项目研究了基因表达变化是否有助于解释这种保护作用。使用聚合酶链反应阵列检查梨状皮层中的炎症细胞因子和受体的表达变化。通过定量聚合酶链反应检查海马中参与脑修复的即时早期基因(Bdnf)、增加神经毒性(Fos)和细胞凋亡控制(Jdp2、Bcl2l1、Bcl2l11)的表达。在梨状皮层中,NIMP 显著刺激了巨噬细胞炎症蛋白 CCL4、IL-1A 和 IL-1B 的表达。肟 20 本身引起的变化最大。当它在 NIMP 后进行治疗时,发生了最大的变化:炎症细胞因子 CCL12 的表达被抑制了 310 倍。在海马中,NIMP 增加了神经毒性标志物 Fos 的表达,并降低了神经保护 Bdnf 和抗细胞凋亡 Bcl2l1 的表达。与 2-PAM 相比,Oxime 20 更刺激了 Bcl2l1 的表达,并使表达更接近载体对照值。