大鼠视网膜急性缺血损伤时视网膜神经节细胞脑红蛋白的表达

发布时间：2018-09-11 07:07

【摘要】： 目的：研究大鼠视网膜急性缺血中视网膜节细胞(Retinal ganglion cells, RGCs)死亡时脑红蛋白(Neuroglobin, Ngb)的表达。 方法：实验研究。成年SD大鼠40只,2-3月龄,体重220-240g,雌性,采用特异性结扎大鼠视网膜动脉的方法造成视网膜急性缺血模型,按照结扎时间不同(0、15、30、60 min)分为4组,每组10只大鼠,均以左眼为实验眼。每组随机选取3只大鼠视网膜用于免疫印迹法检测,取4只大鼠的视网膜分别做荧光金(Fluorogold, FG)逆行标记节细胞计数及免疫组化染色荧光强度分析,3只大鼠做免疫电子显微镜观察。所有数据均采用单因素方差分析进行统计学比较,采用LSD-t检验进行两两比较。 结果：视网膜完全结扎15min时,视网膜中Ngb表达最高(P=0.000),之后逐渐降低,到结扎30min时,Ngb表达接近正常(P=0.728),此时与0min组比较RGCs数量开始减少(P=0.011),结扎60min时,Ngb的表达已明显低于Omin组(P=0.001),与Omin组比较RGCs数量进一步减少(P=0.000)。在视网膜各层中,Ngb的表达以RGCs层为最高,其次为内丛状层与外丛状层。RGCs中Ngb主要分布在胞质,结扎30min时线粒体外室和线粒体嵴也发现有Ngb表达,而内核层及外核层细胞的胞质内始终未见Ngb的表达。 结论：Ngb在大鼠RGCs急性缺血死亡时表达快速升高,主要表达于RGCs的胞质内,与RGCs的生存状态关系密切。
[Abstract]:Aim: to study the expression of (Neuroglobin, Ngb) in rat retinal ganglion cells (RGCs) during (Retinal ganglion cells, RGCs) death in acute retinal ischemia. Methods: experimental study. 40 adult SD rats, weighing 220-240 g and weighing 220-240 g, were divided into 4 groups according to different ligation time (0 ~ 153030 ~ 60 min), each group of 10 rats with left eye as experimental eyes. The retina of each group was randomly selected for immunoblotting assay. The retinal cells of 4 rats were stained with fluorescent gold (Fluorogold, FG) retrograde labeled ganglion cells, and the fluorescence intensity of immunohistochemical staining was analyzed. 3 rats were observed by immunoelectron microscopy. All data were compared by single factor ANOVA and LSD-t test. Results: when the retina was completely ligated with 15min, the expression of Ngb in the retina was the highest (P0. 000), and then decreased gradually. By the time of ligation of 30min, the expression of NGB was close to normal (Pf0.728), and the number of RGCs began to decrease (P0. 011) compared with 0min group. The expression of NGB in 60min ligation group was significantly lower than that in Omin group (P0. 001), and the number of RGCs in Omin group was further decreased compared with Omin group (P0. 000). In all layers of retina, the expression of NGB was highest in RGCs layer, followed by Ngb in inner plexiform layer and outer plexiform layer. Ngb expression was also found in mitochondria outer chamber and mitochondrial crest during ligation of 30min. However, no expression of Ngb was found in the cytoplasm of the inner and outer nuclear layer. Conclusion the expression of% NGB was rapidly increased during the acute ischemic death of RGCs in rats, mainly in the cytoplasm of RGCs, which was closely related to the survival state of RGCs.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1

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