弥漫性轴索损伤后胶质细胞的反应性变化

doi:10.16098/j.issn.0529-1356.2019.05.002

摘要/Abstract

摘要：

目的探讨大鼠弥漫性轴索损伤（DAI）后胶质细胞的反应性变化规律及与轴索继发损伤的相互关系。方法 SD 成年大鼠随机分为对照组及DAI损伤1d、2d、3d、7d组，每组8只。参照Marmarou方法制做 DAI 模型，并对大鼠的脑干部位进行胶质纤维酸性蛋白（GFAP）、离子钙结合适配器分子1（Iba1）、少突胶质细胞系转录因子2（Olig 2）、CC-1、NG2免疫组织化学染色及TUNEL染色，并且通过透射电子显微镜进一步观察脑干超微结构变化。结果 DAI大鼠损伤后3d脑干Iba1标记的小胶质细胞细胞数目显著增多，一直持续至伤后7d，且激活的小胶质细胞呈肥大样形态学改变；DAI大鼠伤后1周内脑干GFAP标记的星形胶质细胞数目虽有所增多，但并没有统计学意义；CC-1标记的成熟少突胶质细胞在DAI后1d即开始明显降低，且随损伤时间的延长而进一步降低。DAI大鼠脑干TUNEL标记的凋亡细胞随损伤时间延长持续增多，与成熟少突胶质细胞的丢失成正相关。Olig2标记的少突胶质细胞系表达在DAI损伤后1周内各时间点均明显增高，于损伤后3d达峰值。NG2标记的少突胶质细胞前体细胞数量于损伤后3d和7d显著增多。透射电子显微镜结果示，DAI大鼠损伤呈现由髓鞘到轴索的特点，最早表现为髓鞘松解，轴索完整；且脱髓鞘会进一步促进轴索骨架崩解，最终导致轴索变性。结论成熟少突胶质细胞对DAI损伤具有选择易感性，髓鞘脱失是轴索继发损伤的重要因素。少突胶质细胞前体细胞增殖，并伴随小胶质细胞的激活。探索胶质细胞的动态变化将为进一步解释轴索继发损伤的病理生理学机制奠定基础。

关键词: 弥漫性轴索损伤, 胶质细胞, 脱髓鞘, 轴索继发损伤, 免疫组织化学, 大鼠

Abstract:

Objective To explore the glial response and the relationship with secondary axonal degeneration in rats after diffuse axonal injury (DAI). Methods Adult male Sprague-Dawley rats were randomly assigned to control or DAI groups sacrificed at 1, 2, 3, 5 and 7 days with 10 rats in each group. DAI model was made referring to modified Marmarou method and glial fibrillary acidic protein(GFAP), ionized calcium binding adaptor molecule-1 (Iba1), recombinant oligodendrocyte lineage transcription factor 2 (Olig2), CC-1, NG2 immunohistochemistry, TUNEL staining and transmission electron microscopy were performed in brain stem.
Results The number of Iba1 labeled positive cells was significantly increased at day 3 and day 7 after injury. Moreover, different hypertrophic morphology was identified after injury. There was no effect of DAI on GFAP expression in brain stem. Numbers of mature oligodendrocyte marker CC-1 immunoreactivity cells within brain stem were significantly decreased at each of the time points after injury. The number of TUNEL positive cells in brain stem was significantly increased with injured time. Olig2 expression was significantly increased throughout the first week and reached peak at day 3 after injury in brain stem. The number of NG2 labeled positive cells was significantly increased at day 3 and day 7 after DAI. Ultrastructural evidence showed myelin release then further developed as widespread delamination and collapse, and leading to degeneration of axonal partner. Conclusion Mature oligodendrocytes are vulnerable in DAI and myelin loss may contribute to axonal degeneration. OPCs proliferate with activation of microglia. This insight of glial response will further explain the pathophysiological mechanism of secondary axonal damage in DAI.

Key words: Diffuse axonal injury, Glial cells, Demyelination, Secondary axonal degeneration, Immunohistochemistry, Rat

李美玉穆娇王婷婷李伟民张国徽 . 弥漫性轴索损伤后胶质细胞的反应性变化[J]. 解剖学报, 2019, 50(5): 554-560.

LI Mei-yu MU Jiao WANG Ting-ting LI Wei-min ZHANG Guo-hui. Glial response following diffuse axonal injury[J]. Acta Anatomica Sinica, 2019, 50(5): 554-560.

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