慢性高眼压状态下小鼠AQP4基因对视网膜胶质细胞活化的影响

发布时间：2018-06-17 12:39

本文选题：慢性高眼压 + 小鼠　；参考：《南京医科大学》2012年硕士论文

【摘要】：目的：研究慢性高眼压状态下水通道蛋白4（aquaporin4,AQP4）基因是否可以通过影响胶质细胞的活化造成青光眼视网膜的损伤，并探讨其可能机制。方法：采用小鼠巩膜外静脉烧烙法建立小鼠高眼压模型。icare回弹式眼压计测量小鼠眼压。右眼为慢性高眼压眼，左眼为对照眼。选择造模成功的慢性高眼压雄性AQP4基因敲除小鼠及其背景雄性野生型小鼠各40只，根据慢性高眼压模型建立的时间（手术结束时开始计算），将两种鼠分别分为5组（24小时、3天、1周、2周、4周），每组8只。石蜡切片行AQP4基因敲除（AQP4-/-）小鼠及野生型（WT）小鼠胶质细胞中胶质纤维酸性蛋白(glial fibrillary acid protein,GFAP)和WT小鼠AQP4的免疫组织化学染色，以及HE染色检测视网膜厚度的变化，荧光显微镜下采集图像。小鼠眼压的组间比较所用统计学方法为t检验、方差分析及SNK法。结果：小鼠巩膜外静脉烧烙术后24小时、3天、1周、2周、4周，实验组眼压与对照组相比差异有统计学意义（P0.05）；HE染色显示：两种鼠在眼压升高后3天视网膜即开始增厚，高眼压1周视网膜明显增厚节细胞水肿，胞浆空亮，内层视网膜细胞间隙增大2周时仍比正常对照组增厚，高眼压4周以后视网膜逐渐萎缩，，可见到部分节细胞胞核变小深染。术后24小时两种鼠GFAP表达开始增加，1周时表达最为明显，2周时开始减弱，4周时最弱，但仍高于对照组。1周时WT小鼠较AQP4基因敲除小鼠GFAP表达增加更为明显。WT小鼠在眼压升高1周时AQP4表达较对照组明显增加，2周、4周时仍高于对照组。WT小鼠在眼压升高1周、2周、4周时AQP4表达与GFAP表达均具有一致性；结论：小鼠巩膜外静脉烧烙的方法能有效的升高小鼠眼压；高眼压状态下AQP4基因可能通过影响小鼠胶质细胞的活化造成青光眼视网膜损伤，其可能作为治疗青光眼的新靶点。
[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.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R774.1

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