谷氨酰胺在大鼠视网膜兴奋性损伤中对HSP70的诱导表达

发布时间：2018-01-19 01:12

本文关键词： 谷氨酰胺热休克蛋白70 N-甲基天冬氨酸青光眼　出处：《青岛大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：在NMDA所致大鼠视网膜兴奋损伤模型中,检测谷氨酰胺诱导HSP70的表达及随时间的变化。方法：健康清洁型Wistar大鼠70只随机分为三组,正常对照组10只,空白对照组10只、实验组即谷氨酰胺腹腔注射组50只。正常对照组不做任何处理,空白对照组和实验组中所有大鼠均随机取大鼠一只眼球玻璃体腔注射2μ l80mmol/lNMDA制作视网膜兴奋性损伤模型,实验组在大鼠视网膜损伤兴奋性损伤模型制作2天后分别腹腔注射谷氨酰胺(0.75g/Kg),并于注射后1h,4h,8h,12h,24h各随机处死10只大鼠,其中1h时同时将空白对照组及正常对照组所有大鼠处死。HE染色光镜下观察各组视网膜组织的形态学变化,免疫组织化学染色方法及Western Blot分析视网膜HSP70表达的变化。结果：HE染色显示正常对照组大鼠视网膜组织结构层次清楚,染色均匀,细胞形态规则。空白对照组与实验组中不同时间段的大鼠视网膜均显示视网膜神经节细胞数目较正常大鼠减少,排列紊乱,神经纤维层与内核层可见萎缩,视网膜整体变薄。Western Blot分析结果显示,正常对照组和空白对照组仅有少量HSP70的表达。实验组注射1h后HSP70的表达开始增强,4h后表达明显增多,8h达到高峰并持续到12h,24h表达较前减少,但仍显著高于对照组。免疫组织化学染色显示诱导表达的HSP70在视网膜各层细胞均有显著表达,而正常对照组及空白对照组HSP70仅微量表达于外核层及光感受器内节。免疫组化染色阳性细胞光密度值测定结果与Westefn Blot结果相吻合。结论：1.成功建立大鼠视网膜兴奋性损伤模型； 2.正常情况下和玻璃体腔注射NMDA2天后大鼠视网膜微量表达HSP70； 3.腹腔注射谷氨酰胺可以在大鼠视网膜兴奋性损伤中诱导HSP70的强烈表达。
[Abstract]:Aim: to detect the expression of glutamine-induced HSP70 and its changes over time in the rat model of retinal excitatory injury induced by NMDA. Methods: 70 healthy and clean Wistar rats were randomly divided into three groups: normal control group (n = 10) and blank control group (n = 10). 50 rats in the experimental group were intraperitoneally injected with glutamine, and no treatment was given in the normal control group. All the rats in the blank control group and the experimental group were randomly selected to inject 2 渭 l 80 mmol / L NMDA into the vitreous body to make the model of retinal excitatory injury. In the experimental group, the excitatory injury model of rat retina was induced by intraperitoneal injection of Glutamine 0.75g / kg / kg respectively 2 days later, and the rats were injected with Glutamine 0.75g / kg / kg for 12 h at 4 h and 8 h after injection respectively. Ten rats were killed at random for 24 hours. At 1 h, all the rats in the blank control group and the normal control group were killed. The morphological changes of retinal tissue in each group were observed under light microscope with HE staining. Immunohistochemical staining and Western Blot were used to detect the expression of HSP70 in retina. Results the histological structure of retina in normal control group was clear and stained evenly. The number of retinal ganglion cells decreased and the number of retinal ganglion cells was disordered and atrophy of nerve fiber layer and nuclear layer were observed in the control group and experimental group. The results of retina thinning. Western Blot analysis showed. There was only a small amount of HSP70 expression in the normal control group and the blank control group, and the expression of HSP70 began to increase at 1 h after injection in the experimental group. The expression of HSP70 increased significantly at 8 h after injection and reached its peak at 8 h and lasted until 12 h. The expression of HSP70 was significantly decreased in 24 hours, but still significantly higher than that in the control group. Immunohistochemical staining showed that the expression of HSP70 was significantly expressed in all layers of the retina. However, HSP70 was only expressed in the outer nuclear layer and the inner ganglia of the photoreceptor in the normal control group and the blank control group. The results of optical density of the positive cells by immunohistochemical staining and Westefn were compared with those of the normal control group and the blank control group. The Blot results were in agreement with each other. Conclusion 1. The rat model of retinal excitatory injury was established successfully. 2. Normal and intravitreal injection of HSP70 in the retina of rats; 3.Intraperitoneal injection of glutamine could induce the strong expression of HSP70 in the excitatory injury of rat retina.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R774.1

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