A study of neurogenic bone lesion in neuropathic pain model (2);using immobilized model mouse and osteopetrosis mouse

神经性疼痛的病因有许多不确定的地方。此外，神经性疼痛有时还伴有骨骼萎缩。我们用酶免疫化学方法检查骨萎缩。此外，我们还通过行为学、生物化学和免疫组织化学的方法研究了巨噬细胞、肿瘤坏死因子-α与热痛敏的关系。用TRAP染色和免疫组织化学方法分别对破骨细胞和P物质(SP)进行染色。然后计算每区破骨细胞的数量。在后期的检查中，我们制作了正常小鼠(对照组)和骨化病(OP/OP)小鼠的神经痛模型，其巨噬细胞先天缺失。观察各组大鼠戒断阈值时间、免疫印迹法检测肿瘤坏死因子-α、免疫组织化学检测巨噬细胞和肿瘤坏死因子-α的变化。但三组间差异无统计学意义。在SP免疫反应中，CCI和IC均可见深染区，但组间比较差异无统计学意义。在后一种检查中，在12h后，对照组出现热痛觉过敏，而OP/OP组未见痛觉过敏，巨噬细胞和肿瘤坏死因子-α的表达明显低于对照组。伤后5天，两组动物均出现热痛敏反应，12小时后巨噬细胞和肿瘤坏死因子-α表达相似，破骨细胞表达上调可能是除制动外的其他因素所致。巨噬细胞和肿瘤坏死因子-α可能在神经损伤后早期与热痛觉过敏有关，但随着时间的推移可能与热痛觉过敏无关。有必要研究骨萎缩与SP以外的神经肽及中枢神经系统改变的关系。

Neuropathic pain has many uncertain points of the cause. In addition, neuropathic pain is sometimes accompanied by bone atrophy. We examined bone atrophy by using enzymeimmunochemistry. Additionally, we also examined the relationships between macrophages, tumor necrosis factor (TNF)-α and thermal hyperalgesia by using behavioristy, biochemistry and immunohistochemistry.In former examination, we made neuropathic pain (CCI), immobilized (IMO) and immobilized with neuropathic pain model (IC). Then, we stained osteoclasts and substance P (SP) by TRAP staining and immunohistochemistry, respectively. Then, we calculated osteoclasts number per area.In latter examination, we made neuropahtic pain model to normal mouse (control) and osteopetrosis (op/op) mouse, whose macrophages are absent congenitally. Then, we measured withdrawal thresholds time, analyzed TNF-α by western blotting, and observed macrophages and TNF-α by immunohistochemistry.Osteoclasts number was larger in ipsilateral side than in contralateral side in CCI and IC at post-1 week, and in IC at post-3 weeks. However, they were not significantly differences between three groups. In SP immunoreactivity, deep staining areas were observed in CCI and IC, but there were not significantly different between these groups. In latter examination, at post-12 hours, thermal hyperalgesia was observed in control but not op/op, and expression of macrophages and TNF-α was fewer in op/op than in control. At post-5 days, thermal hyperalgesia was observed in both, but expression of macrophages and TNF-α was similar in post 12 hours.Upregulation of osteoclasts in CCI may be occurred by some factors other than immobilization. There are possibilities that macrophages and TNF-α may relate to thermal hyperalgesia at early phase after nerve lesion, but may not relate to thermal hyperalgesia as time passed. It is necessary to research relations between bone atrophy and neuropeptides other than SP, and change of central nervous system.