本文關鍵詞： 谷氨酰胺 熱休克蛋白70 N-甲基天冬氨酸 青光眼　出處：《青島大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的：在NMDA所致大鼠視網(wǎng)膜興奮損傷模型中,檢測谷氨酰胺誘導HSP70的表達及隨時間的變化。 方法：健康清潔型Wistar大鼠70只隨機分為三組,正常對照組10只,空白對照組10只、實驗組即谷氨酰胺腹腔注射組50只。正常對照組不做任何處理,空白對照組和實驗組中所有大鼠均隨機取大鼠一只眼球玻璃體腔注射2μ l80mmol/lNMDA制作視網(wǎng)膜興奮性損傷模型,實驗組在大鼠視網(wǎng)膜損傷興奮性損傷模型制作2天后分別腹腔注射谷氨酰胺(0.75g/Kg),并于注射后1h,4h,8h,12h,24h各隨機處死10只大鼠,其中1h時同時將空白對照組及正常對照組所有大鼠處死。HE染色光鏡下觀察各組視網(wǎng)膜組織的形態(tài)學變化,免疫組織化學染色方法及Western Blot分析視網(wǎng)膜HSP70表達的變化。 結果：HE染色顯示正常對照組大鼠視網(wǎng)膜組織結構層次清楚,染色均勻,細胞形態(tài)規(guī)則�？瞻讓φ战M與實驗組中不同時間段的大鼠視網(wǎng)膜均顯示視網(wǎng)膜神經節(jié)細胞數(shù)目較正常大鼠減少,排列紊亂,神經纖維層與內核層可見萎縮,視網(wǎng)膜整體變薄。Western Blot分析結果顯示,正常對照組和空白對照組僅有少量HSP70的表達。實驗組注射1h后HSP70的表達開始增強,4h后表達明顯增多,8h達到高峰并持續(xù)到12h,24h表達較前減少,但仍顯著高于對照組。免疫組織化學染色顯示誘導表達的HSP70在視網(wǎng)膜各層細胞均有顯著表達,而正常對照組及空白對照組HSP70僅微量表達于外核層及光感受器內節(jié)。免疫組化染色陽性細胞光密度值測定結果與Westefn Blot結果相吻合。 結論：1.成功建立大鼠視網(wǎng)膜興奮性損傷模型； 2.正常情況下和玻璃體腔注射NMDA2天后大鼠視網(wǎng)膜微量表達HSP70； 3.腹腔注射谷氨酰胺可以在大鼠視網(wǎng)膜興奮性損傷中誘導HSP70的強烈表達。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R774.1

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