实验性脊髓损伤后胶质瘢痕形成病理学规律与影像学特征

发布时间：2018-05-04 18:43

本文选题：实验性脊髓损伤 + 胶质瘢痕　；参考：《第三军医大学》2005年博士论文

【摘要】： 脊髓损伤(spinal cord injury, SCI)后,胶质瘢痕(glial scar)的形成是慢性SCI的特征之一,并且是脊髓损伤后神经再生障碍的重要原因,脊髓损伤如脊髓挫伤后反应性胶质化(reactive gliosis),是中枢神经系统损伤后慢性期的主要病变之一,这种反应性胶质化,将坏死组织与脊髓隔离,并使前者液化形成囊腔,一方面阻止脊髓的继发性损害,另一方面也阻碍了轴突再生。如何在移植治疗中判断切除胶质瘢痕的时机与范围,是神经外科的难题之一。以往的研究对胶质瘢痕的空间分布特征缺乏研究,目前也无切除胶质瘢痕的适宜方法。本研究拟制备打击伤SCI模型,观察SCI的影像学与病理学表现及大鼠慢性SCI胶质瘢痕的时间与空间分布,希望能发现胶质瘢痕分布规律,为彻底去除胶质瘢痕提供形态学依据。材料与方法 1.采用犬为脊髓打击伤模型的损伤能量为30g×15cm,重点对SCI后组织学与影像学变化对比观察,观察SCI影像学变化的病理基础;采用SD大鼠为脊髓打击伤模型的损伤能量为30g×5cm,以组织学、神经运动功能评分、诱发电位测试、免疫组织化学、免疫荧光组织化学双标记、BDA顺行神经纤维示踪、TMR与HRP逆行神经纤维示踪等方法进行观察,且大鼠所有标本均采用恒冷箱矢状纵行切片,每个脊髓标本切片100张,脊髓全部标本10套均进行不同染色观察,每张载玻上每两张切片间隔200μm,每张切片均能观察到损伤区及损伤区两端5mm内残留组织,经普通光学微镜及荧光显微镜下观察并进行图像拼接,观察损伤区胶质瘢痕分布的形成时间与空间分布规律,胶质瘢痕厚度及胶质瘢痕区轴突与星形胶质细胞的空间关系。 2.采用免疫组织化学染色观察SCI后神经干细胞标志物nestin与轴突生长抑制因子nogo在SCI后不同时间、不同部位的表达、分布,并分别与NF免疫荧光双标观察SCI后损伤区范围神经再生内在修复能力变化规律,轴突生长抑制因子nogo在损伤区分布及其与轴突再生的相互关系。 3.脊髓损伤大鼠在伤后8周采用BDA顺行神经纤维示踪、TMR与HRP逆行神经纤维示踪结合免疫荧光组织化学、NF与GFAP荧光双标方法,在矢状纵切片不同部
[Abstract]:After spinal cord injury, glial scar formation is one of the characteristics of chronic SCI, and it is an important cause of nerve regeneration disorder after spinal cord injury. Spinal cord injury, such as reactive gliosism after spinal cord contusion, is one of the main pathological changes in the chronic stage after central nervous system injury, which separates necrotic tissue from spinal cord and liquefies the former into cystic cavity. On the one hand to prevent secondary damage to the spinal cord, on the other hand also hindered axonal regeneration. How to judge the time and scope of excision of glial scar in transplantation is one of the difficult problems in neurosurgery. Previous research on the spatial distribution of glial scar is lack of research, and there is no suitable method to remove glial scar. The aim of this study was to establish a SCI model of striking injury and to observe the imaging and pathological features of SCI and the temporal and spatial distribution of chronic SCI glial scar in order to find the regularity of glial scar distribution and provide morphological basis for removing glial scar completely. Materials and methods 1. The injury energy of spinal cord injury model in dogs was 30g 脳 15cm. The histopathological basis of SCI was observed by comparing the histological and imaging changes after SCI, and the injury energy of SD rats was 30g 脳 5cm for histology. The neuromotor function score, evoked potential test, immunohistochemistry, immunofluorescence histochemical double labeling, TMR and HRP retrograde nerve fiber tracing were observed. Sagittal longitudinal sections were used in all the specimens of rats. 100 spinal cord sections and 10 spinal cord specimens were observed with different staining. Every two slices on each glass were separated by 200 渭 m, and the residual tissues in the injured area and the two ends of the injured area were observed in each slice. The residual tissues were observed under ordinary optical microscopes and fluorescence microscope, and the images were spliced together. The formation time and spatial distribution of glial scar, the thickness of glial scar and the spatial relationship between axons and astrocytes were observed. 2. Immunohistochemical staining was used to observe the expression and distribution of neural stem cell marker nestin and axon growth inhibitor nogo at different time points after SCI. The changes of nerve regeneration ability and the distribution of axon growth inhibitory factor (nogo) in the injured area after SCI were observed with NF immunofluorescence double labeling, and the relationship between nogo and axon regeneration was also observed. 3. Eight weeks after spinal cord injury, BDA anterograde nerve fiber tracer and HRP retrograde nerve fiber tracing combined with immunofluorescence histochemical staining (NF) and GFAP fluorescence double labeling method were used in the sagittal longitudinal sections of rats.
【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R361

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