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本实验旨在建立可用于中枢神经系统Waller变性研究的大鼠视神经(ON)损伤模型。成年SD大鼠12只,用夹持力为148g、镊端宽1mm的小号动脉阻血镊于球后2mm处夹持左侧视神经,致伤时间10s。术后随机分为两组(每组6只):一组动物术后立即于双侧上丘表面放置逆行荧光染料荧光金(FG),72h后处死。取视网膜铺片,ON冰冻连续切片。另一组动物术后72h常规灌注固定,取ON冰冻连续切片,分别行β-tubulin及NF-160免疫组化染色和HE染色。结果显示:左侧ON夹伤后3d,同侧视网膜内未见FG逆行标记的视网膜神经节细胞,ON夹伤部位形成一条宽达1mm的损伤带,ON基质连续,FG标记的神经纤维被阻断于损伤远侧,损伤远侧的β-tublin阳性纤维多已崩溃,NF-160阳性纤维数量较多,部分纤维末梢形成膨大的回缩球。上述结果表明ON夹伤后神经元轴突完全离断,中枢神经基质的连续性得以保持,损伤远侧段轴突发生典型的Waller变性,从而为中枢神经Waller变性及其药物治疗研究的筛选和评估提供了简单实用的实验模型。
The aim of this study was to establish a rat optic nerve (ON) injury model that could be used to study Waller degeneration in central nervous system. 12 adult Sprague-Dawley rats were used to clamp the left optic nerve at a distance of 2mm posterior to the trochanter with a clamping force of 148g and a tweezers end with a width of 1mm. The injury time was 10s. All patients were randomly divided into two groups (6 in each group): One group of animals placed fluorescent dye gold (FG) retrograde fluorescent dye on the surface of the bilateral superior colliculus immediately after operation, and were sacrificed after 72 hours. Take retinal film, ON frozen serial slices. The other group of animals were routinely perfused and fixed at 72 hours after operation, and ON frozen serial sections were taken for immunohistochemical staining of β-tubulin and NF-160 and HE staining respectively. The results showed that retinal ganglion cells with retrograde retrograde index were not found in the ipsilateral retinal injury on the left side of ON injury. Injured retinal ganglion cells with a width of 1 mm were formed at the site of ON injury. The ON matrix was continuous and the FG-labeled nerve fibers were blocked Broken distal to the injury, the distal β-tublin-positive fibers have collapsed, NF-160-positive fibers more number of part of the formation of the distal tip of the bulging retractable ball. The above results show that the neuronal axon completely breaks down after ON clip, the continuity of the central nervous system matrix is maintained, and the typical Waller’s degeneration of the distal axonal is impaired so as to screen for Waller’s degeneration of central nervous system and its drug treatment And evaluation provides a simple and practical experimental model.