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点。此外，神经性疼痛有时还伴有骨萎缩。我们通过使用酶免疫化学检查骨萎缩。此外，我们还利用行为学、生物化学和免疫组织化学研究了巨噬细胞、肿瘤坏死因子(TNF)-α与热痛觉过敏之间的关系。在前期的研究中，我们制作了神经病理性疼痛(CCI)、固定化(IMO)和固定化神经病理性疼痛模型(IC)。然后，我们分别通过TRAP染色和免疫组织化学对破骨细胞和P物质（SP）进行染色。然后，我们计算每个区域的破骨细胞数量。在后期检查中，我们对正常小鼠（对照）和先天性缺乏巨噬细胞的骨石症（op/op）小鼠制作神经性疼痛模型。然后测量戒断阈值时间，用Western blotting分析TNF-α，并用免疫组织化学观察巨噬细胞和TNF-α。CCI和IC后1周、IC后3周破骨细胞数同侧破骨细胞数量多于对侧。然而，三组之间没有显着差异。在SP免疫反应中，CCI和IC中观察到深染色区域，但这些组之间没有显着差异。在后一次检查中，12小时后，在对照中观察到热痛觉过敏，但在op/op中未观察到，并且op/op中巨噬细胞和TNF-α的表达比对照少。 5天后，两者均观察到热痛觉过敏，但12小时后巨噬细胞和TNF-α的表达相似。CCI中破骨细胞的上调可能是由固定以外的其他因素引起的。巨噬细胞和TNF-α可能与神经损伤后早期的热痛觉过敏有关，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、中枢神经系统变化的关系。

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.