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目的:周围神经再生过程中巨噬细胞发挥了重要的作用,然而目前对于神经内内源性和外源性巨噬细胞的具体作用了解的却很少,因此本实验研究了小鼠坐骨神经损伤后早期再生过程中内源性和外源性巨噬细胞数量比例变化的情况,探索周围神经再生的规律。方法:移植CAG-EGFP转基因小鼠的全骨髓有核细胞到骨髓灭活野生型C57Bl/6小鼠体内建立嵌合体小鼠模型。待移植成功3个月后夹伤小鼠一侧坐骨神经,并在损伤后第2、7、14和28天取材、切片,使用巨噬细胞特异性抗体CD68进行免疫荧光染色,分析损伤神经段中内源性巨噬细胞(CD68+/EGFP-)、外源性巨噬细胞(CD68+/EGFP+)的数量及其比例变化情况。结果:①夹伤骨髓移植模型小鼠坐骨神经后,参与坐骨神经损伤修复的巨噬细胞可分为两类,即内源性巨噬细胞(CD68+/EGFP-)和外源性巨噬细胞(CD68+/EGFP+);②夹伤坐骨神经后,浸润的总巨噬细胞数量从第2天开始逐渐增加,到第14天达到高峰,约为正常情况下的60倍,随后逐渐减少;③起初外、内源性巨噬细胞间的比例是1:1,差值最大出现在损伤后第14天为4:1。结论:小鼠坐骨神经夹伤后,内外源性巨噬细胞共同参与了受损神经组织远心段的修复和再生过程,损伤初期发挥作用的主要是内源性巨噬细胞,随后大量浸润的外源性巨噬细胞占主导作用。本实验首次连续观察并定量分析了神经损伤后早期内源性和外源性巨噬细胞的数量改变,证实了瓦勒氏变性过程中内源性和外源性巨噬细胞在不同阶段对巨噬细胞总量的贡献作用。
OBJECTIVE: Macrophages play an important role in the process of peripheral nerve regeneration. However, little is known about the specific role of endogenous and exogenous macrophages in neurons. Therefore, in this study, after sciatic nerve injury in mice Changes in the proportion of endogenous and exogenous macrophages during early regeneration and explore the rules of peripheral nerve regeneration. METHODS: Chimeric mouse models were established by transplanting whole bone marrow cells of CAG-EGFP transgenic mice into bone marrow-inactivated wild type C57BI / 6 mice. Three months after transplantation, sciatic nerves were injured in one side of the sciatic nerve and harvested on the 2nd, 7th, 14th and 28th day after the injury. The sections were stained and immunofluorescence staining was performed using macrophage-specific antibody CD68 to analyze the damaged nerve segments The changes of the number of endogenous macrophages (CD68 + / EGFP-) and exogenous macrophages (CD68 + / EGFP +) and their proportions. Results: ① Macrophages involved in the repair of sciatic nerve injury can be divided into two groups, namely, endogenous macrophages (CD68 + / EGFP-) and exogenous macrophages (CD68 + / EGFP- EGFP +); ② After sciatic nerve injury, the total number of infiltrating macrophages gradually increased from the second day to the peak on the fourteenth day, which was about 60 times of that in the normal condition, then decreased gradually; ③ At first, The ratio of macrophages was 1: 1, with the largest difference being 4: 1 on day 14 after injury. Conclusion: After sciatic nerve injury in mice, both endogenous and exogenous macrophages participate in the process of repair and regeneration of the distal segment of the damaged nerve tissue. The initial stage of injury is mainly endogenous macrophages, followed by extensive infiltration Derived macrophages play a dominant role. This experiment for the first time continuous observation and quantitative analysis of the early endogenous and exogenous macrophages after nerve injury changes in the number of confirmed Wallerian degeneration during endogenous and exogenous macrophages in different stages of giant The contribution of total macrophages.