本文選題：谷氨酰胺 + 眼挫傷　； 參考：《遼寧醫(yī)學院》2012年碩士論文

【摘要】：目的 探討谷氨酰胺(Glutamine, Gln)對視網膜HSP27（Heat shock protein27）和NF-κB（Nuclear factor-kappa B）的調節(jié)作用，明確Gln對挫傷大鼠視網膜保護作用機制，以期為臨床治療視網膜挫傷提供實驗依據(jù)。 方法 SD雄性大鼠45只，5只用于正常對照組。余下40只用于制備眼挫傷模型。正常組不處理，模型組和Gln組仿Allen’s重擊法制作視網膜挫傷模型致左眼挫傷。模型制備成功后，40只大鼠隨機分為2組，視網膜挫傷模型組(模型組)和Gln處理組(Gln組)。后兩組又分為12h，24h，3天和7天4個時間段，每組每個時間段大鼠5只。傷后第2天，Gln組腹腔注射Gln（0.5g/kg），每日一次，模型組用同體積的生理鹽水腹腔注射。Gln組、模型組各時段于給藥給水停止后第2天取材，所有組別的大鼠均取左眼。應用HE染色方法觀察各組視網膜各層細胞的形態(tài)改變；應用免疫組織化學方法檢測視網膜組織中HSP27和NF-κB蛋白的表達變化。 結果 1、視網膜各層組織病理學改變 HE染色觀察可見模型組與Gln組視網膜全層細胞數(shù)減少，視網膜厚度較正常組明顯變薄，各層細胞排列紊亂，細胞水腫；正常對照組視網膜各層細胞數(shù)量多，排列整齊，形態(tài)較完整。 2、HSP27蛋白檢測結果 光鏡下HSP27蛋白免疫反應產物呈棕黃色。HSP27蛋白在正常組視網膜神經節(jié)細胞層、內核層胞漿內有表達，在外核層和感光細胞細胞層的胞漿內表達極少。損傷12h后，在模型組和Gln干預組，HSP27蛋白在視網膜的表達變化不明顯，與正常組比較，無統(tǒng)計學意義。損傷24h后，模型組HSP27蛋白的表達明顯下降，明顯低于正常組（P0.05）3d后，HSP27蛋白表達降至最低，7d后，其表達有所恢復，但仍高于正常組（P0.05）。在Gln組，HSP27蛋白表達變化趨勢與模型組相似，但在24h、3d、7d, HSP27蛋白表達較模型組明顯增強，其蛋白陽性產物的平均光密度值(Mean optical density, MOD)明顯高于模型組（P0.01）。 3、NF-κB蛋白檢測結果 NF-κB蛋白免疫反應產物呈棕黃色，在神經節(jié)細胞層、內核層和外核層及感光細胞層的胞漿均有表達。正常組大鼠視網膜組織中有適量NF-κB蛋白表達。在模型組，損傷后12h，NF-κB蛋白在視網膜的表達變化不明顯，與正常組比較，無統(tǒng)計學意義。損傷后24h，模型組NF-κB蛋白的表達升高，明顯高于正常組（P0.01）；損傷后3d，NF-κB蛋白表達達峰值，至7d后，其表達略有下降，但仍高于正常組（P0.05）。在Gln組，除12h外，NF-κB蛋白表達在24h、3d、7d均較模型組明顯降低，其蛋白陽性產物的平均光密度值(Mean optical density,MOD)明顯低于模型組（P0.01）。 結論 HSP27和NF-κB可能參與了眼挫傷視網膜病變過程；谷氨酰胺上調了挫傷視網膜神經細胞HSP27蛋白表達，，下調了NF-κB蛋白表達，這可能是谷氨酰胺對眼挫傷視網膜病變產生保護作用的機理之一。
[Abstract]:Purpose To investigate the regulatory effects of glutamine Glutamine27 (Glutamine27) and NF- 魏 B(Nuclear factor-kappa B on retinal contusion in rats, and to clarify the protective mechanism of Gln on retina contusion in order to provide experimental evidence for clinical treatment of retinal contusion. Method 45 SD male rats were used as normal control group. The remaining 40 cases were used to establish the model of eye contusion. The model group and Gln group made retinal contusion model by Allen's method. After the establishment of the model, 40 rats were randomly divided into two groups: retinal contusion model group (model group) and Gln treatment group. The latter two groups were divided into two groups: 12 h, 24 h, 3 days and 7 days, with 5 rats in each group. On the second day after injury, the GLN group was injected intraperitoneally with Glnn 0.5g / kg / kg, once a day. The model group was injected intraperitoneally with the same volume of normal saline. The rats in the model group were collected on the second day after the drug supply stopped, and the left eyes were taken from all groups of rats. The morphological changes of retinal cells in each group were observed by HE staining and the expression of HSP27 and NF- 魏 B in retinal tissues were detected by immunohistochemical method. Result 1. Histopathological changes in all layers of retina He staining showed that the number of whole retinal cells in the model group and Gln group decreased, the thickness of the retina became thinner than that in the normal group, the cells in each layer were disordered and the cells were edema, while in the normal control group, the number of cells in each layer of the retina was more than that in the normal control group. The shape is relatively complete. Detection of HSP27 protein Under light microscope, the immunoreactive product of HSP27 protein was brownish yellow. HSP27 protein was expressed in the retinal ganglion cell layer of normal group, and in the cytoplasm of nuclear layer, but in the outer nuclear layer and photosensitive cell layer. After 12 hours of injury, the expression of HSP27 protein in the retina of the model group and Gln intervention group was not significantly changed, and there was no significant difference between the model group and the normal group. After 24 hours of injury, the expression of HSP27 protein in the model group was significantly lower than that in the normal group. The expression of HSP27 protein in the model group was significantly lower than that in the normal group after 3 days, and the expression of HSP27 protein in the model group was lower than that in the control group. After 7 days of injury, the expression of HSP27 protein recovered, but still higher than that in the normal group. The change trend of HSP27 protein expression in Gln group was similar to that in model group, but the expression of HSP27 protein was significantly higher than that in model group at 24 h or 3 d, and the mean optical density of the protein positive product was significantly higher than that in model group (P 0.01). 3detection of NF- 魏 B protein The immunoreactive products of NF- 魏 B protein were brown and expressed in the cytoplasm of ganglion cell layer, nuclear layer, outer nuclear layer and photosensitive cell layer. In normal group, NF- 魏 B protein was expressed in retina tissue. In the model group, the expression of NF- 魏 B protein in the retina was not significantly changed at 12 h after injury, but there was no statistical significance compared with the normal group. The expression of NF- 魏 B protein in the model group was significantly higher than that in the normal group at 24 h after injury, and reached a peak at 3 days after injury, and decreased slightly at 7 days after injury, but still higher than that in the normal group. In the Gln group, the expression of NF- 魏 B protein was significantly lower than that of the model group on the 3rd day after 24 h, and the mean optical density of the protein positive product was significantly lower than that of the model group (P 0.01). Conclusion HSP27 and NF- 魏 B may be involved in the process of eye contusion retinopathy, glutamine upregulated the expression of HSP27 protein and down-regulated the expression of NF- 魏 B protein in retinal nerve cells. This may be one of the mechanisms of the protective effect of glutamine on ocular contusion retinopathy.
【學位授予單位】：遼寧醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R779.1

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