本文選題：慢性高眼壓 + 小鼠　； 參考：《南京醫(yī)科大學》2012年碩士論文

【摘要】：目的：研究慢性高眼壓狀態(tài)下水通道蛋白4（aquaporin4,AQP4）基因是否可以通過影響膠質細胞的活化造成青光眼視網(wǎng)膜的損傷，并探討其可能機制。 方法：采用小鼠鞏膜外靜脈燒烙法建立小鼠高眼壓模型。icare回彈式眼壓計測量小鼠眼壓。右眼為慢性高眼壓眼，左眼為對照眼。選擇造模成功的慢性高眼壓雄性AQP4基因敲除小鼠及其背景雄性野生型小鼠各40只，根據(jù)慢性高眼壓模型建立的時間（手術結束時開始計算），將兩種鼠分別分為5組（24小時、3天、1周、2周、4周），每組8只。石蠟切片行AQP4基因敲除（AQP4-/-）小鼠及野生型（WT）小鼠膠質細胞中膠質纖維酸性蛋白(glial fibrillary acid protein,GFAP)和WT小鼠AQP4的免疫組織化學染色，以及HE染色檢測視網(wǎng)膜厚度的變化，熒光顯微鏡下采集圖像。小鼠眼壓的組間比較所用統(tǒng)計學方法為t檢驗、方差分析及SNK法。 結果：小鼠鞏膜外靜脈燒烙術后24小時、3天、1周、2周、4周，實驗組眼壓與對照組相比差異有統(tǒng)計學意義（P0.05）；HE染色顯示：兩種鼠在眼壓升高后3天視網(wǎng)膜即開始增厚，高眼壓1周視網(wǎng)膜明顯增厚節(jié)細胞水腫，胞漿空亮，內層視網(wǎng)膜細胞間隙增大2周時仍比正常對照組增厚，高眼壓4周以后視網(wǎng)膜逐漸萎縮，，可見到部分節(jié)細胞胞核變小深染。術后24小時兩種鼠GFAP表達開始增加，1周時表達最為明顯，2周時開始減弱，4周時最弱，但仍高于對照組。1周時WT小鼠較AQP4基因敲除小鼠GFAP表達增加更為明顯。WT小鼠在眼壓升高1周時AQP4表達較對照組明顯增加，2周、4周時仍高于對照組。WT小鼠在眼壓升高1周、2周、4周時AQP4表達與GFAP表達均具有一致性； 結論：小鼠鞏膜外靜脈燒烙的方法能有效的升高小鼠眼壓；高眼壓狀態(tài)下AQP4基因可能通過影響小鼠膠質細胞的活化造成青光眼視網(wǎng)膜損傷，其可能作為治療青光眼的新靶點。
[Abstract]:Aim: to investigate whether the aquaporin 4aquaporin 4 (AQP4) gene can affect the activation of glial cells and to investigate the mechanism of retinal damage induced by chronic intraocular hypertension. Methods: mouse intraocular pressure model was established by scleral vein cauterization. The right eye was chronic high intraocular pressure and the left eye was the control eye. The successful chronic intraocular pressure male AQP4 knockout mice and their background male wild type mice were selected. According to the time of establishment of chronic high intraocular pressure model (starting to calculate at the end of operation), the two kinds of rats were divided into five groups respectively: 5 groups were divided into 5 groups, 8 rats in each group were divided into 5 groups: 24 hours, 3 days, 1 week, 2 weeks and 4 weeks. Immunohistochemical staining of glial fibrillary acidic protein (glial fibrillary acid protein) in glial cells of AQP4 knockout AQP4-r-) mice and wild type WT-mice were performed in paraffin sections, and the changes of retinal thickness were detected by HE staining. The images were collected under fluorescence microscope. The statistical methods of intraocular pressure in mice were t test, ANOVA and SNK. Results: the intraocular pressure in the experimental group was significantly different from that in the control group (P 0.05) and HE staining showed that the retina began to thicken 3 days after intraocular pressure elevation. After 1 week of high intraocular pressure, retinal thickening ganglion cells were edema, cytoplasm was empty, and the inner retinal cell gap was still thicker than that of the normal control group at 2 weeks. After 4 weeks of high intraocular pressure, the retina gradually shrank, and some of the ganglion cell nuclei became small and deep stained. The expression of GFAP in both groups began to increase 24 hours after operation. The expression of GFAP was the most obvious at 1 week and the weakest at 4 weeks. The expression of GFAP in WT mice was significantly higher than that in AQP4 knockout mice at week 1. The expression of AQP4 in WT mice was significantly higher than that in control group at 1 week after intraocular pressure elevation. The expression of AQP4 was consistent with that of GFAP at 2 weeks and 4 weeks. Conclusion: the method of scleral vein cauterization can effectively elevate the intraocular pressure in mice, and AQP4 gene may cause glaucoma retinal injury by affecting the activation of mouse glia cells under high intraocular pressure. It may be a new target for the treatment of glaucoma.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R774.1

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相關期刊論文 前2條

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