NMDA所致大鼠视网膜兴奋性损伤后Brn-3a表达的变化
发布时间：2018-12-13 19:46
【摘要】：目的：研究NMDA(N-methyl-D-aspartate, N-甲基-D-天门冬氨酸)所致大鼠视网膜兴奋性损伤后Brn3a表达的变化。 方法：健康成年清洁级SD大鼠30只,随机分为正常对照组(6只),实验组(24只)。实验组所有大鼠右眼玻璃体腔注射NMDA作为实验眼,左眼玻璃体腔注射PBS作为实验对照眼,随机分成A, B, C, D四个组(每组6只)分别在造模后8h,16h,24h,48h处死,免疫组织化学法观察视网膜Brn3a表达的变化,实时定量RT-PCR法观察视网膜Brn3a mRNA的变化。 结果：1光学显微镜：实验对照眼和正常眼视网膜组织结构层次清楚,染色均匀,细胞形态规整。实验眼在造模后8h,部分视网膜神经节细胞(RGCs)内出现空泡变性；造模后16h,视网膜神经节细胞(RGCs)数目较正常眼减少,排列疏松,部分细胞出现核固缩,胞内空泡变性逐渐明显,视网膜内核层变薄；造模后24h,视网膜神经节细胞明显减少,核固缩,核周空泡样变性明显,视网膜内核层细胞排列疏松紊乱,厚度变薄；造模后48h,视网膜神经节细胞已较少,偶尔可见,核固缩,浓染,胞内空泡样变性,细胞形态不规则,视网膜内核层结构更加紊乱,厚度更薄。2、免疫组织化学：Brn3a只在视网膜神经节细胞层中表达,在实验眼,Brn3a在造模后8h表达与正常视网膜相比变化不明显,造模16h后,Brn3a+RGCs已有减少,24h后,Brn3a+RGCs数目明显减少,48h后,Brn3a+RGCs分布稀疏,零星可见。实验对照组Brn3a+RGCs较空白组Brn3a+RGCs表达无明显差别。3、实时定量PCR：实验组Brn3amRNA表达在造模后8h较正常对照组减少(P0.05),为正常对照组的78.7%。在16h后明显减少,为正常对照组的27.1%,24h后表达量为正常对照组的11.5%,48h表达继续降低,实际表达量仅为正常对照组的6.2%。而实验对照眼与空白对照眼视网膜中Brn3amRNA的表达量无明显差别(P0.05)。 结论：NMDA可造成大鼠视网膜兴奋性损伤,导致视网膜神经节细胞数目减少,视网膜中Brn3a及其mRNA的表达减少。
[Abstract]:Aim: to study the changes of Brn3a expression in rat retina excitatory injury induced by N-methyl-D-aspartate (N-methyl-D-aspartate). Methods: 30 healthy adult SD rats were randomly divided into control group (n = 6) and experimental group (n = 24). All rats in the experimental group were injected with NMDA into the vitreous cavity of the right eye and PBS as the experimental control eye in the left eye. They were randomly divided into four groups of A, B, C, D (6 rats in each group). The rats were sacrificed at 8 h, 16 h, 24 h and 48 h, respectively. The changes of Brn3a expression in retina were observed by immunohistochemical method, and the changes of Brn3a mRNA in retina were observed by real time quantitative RT-PCR. Results: 1 Optical microscope: the retinal tissue structure of the experimental control eyes and normal eyes were clear, stained evenly, and the morphology of the cells was regular. Vacuolar degeneration occurred in some retinal ganglion cells (RGCs) at 8 hours after the model. After 16 hours, the number of (RGCs) in retinal ganglion cells decreased compared with the normal eyes, the arrangement was loose, some cells showed nuclear shrinkage, the degeneration of intracellular vacuoles became obvious, and the inner layer of retina became thinner. After 24 hours, the retinal ganglion cells decreased obviously, the nucleus became pyknosis, the degeneration of perinuclear vacuole was obvious, the arrangement of retinal nuclear layer cells was loose and disordered, and the thickness of retinal nuclear layer became thinner. 48 hours after the model, the retinal ganglion cells were few, occasionally visible, nuclear pyknosis, dense staining, vacuolar degeneration, irregular morphology, the structure of the retinal nuclear layer is more disorder, thickness is thinner. 2. Immunohistochemical staining: Brn3a was only expressed in the retinal ganglion cell layer. In experimental eyes, the expression of Brn3a was not significantly different from that of normal retina at 8 h after modeling. After 16 h of modeling, the expression of Brn3a RGCs had been reduced, and the number of Brn3a RGCs was significantly decreased after 24 h, and 48 h later. Brn3a RGCs distribution is sparse and sporadic. Compared with the control group, the expression of Brn3a RGCs in the experimental group was not significantly different from that in the blank group. 3. The expression of Brn3amRNA in the real-time quantitative PCR: group was significantly lower than that in the normal control group at 8 h after the establishment of the model (P0.05), which was 78.7% of the normal control group. After 16 hours, the expression level of 27.1g / 24 h in the normal control group was lower than that in the normal control group (11.5h / 48h), but the actual expression level was only 6.2% of the normal control group (P < 0.05). However, there was no significant difference in the expression of Brn3amRNA between the experimental control eyes and the blank control eyes (P0.05). Conclusion: NMDA can induce excitatory injury of rat retina, decrease the number of retinal ganglion cells and decrease the expression of Brn3a and mRNA in the retina.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R779.1
本文编号：2377111
[Abstract]:Aim: to study the changes of Brn3a expression in rat retina excitatory injury induced by N-methyl-D-aspartate (N-methyl-D-aspartate). Methods: 30 healthy adult SD rats were randomly divided into control group (n = 6) and experimental group (n = 24). All rats in the experimental group were injected with NMDA into the vitreous cavity of the right eye and PBS as the experimental control eye in the left eye. They were randomly divided into four groups of A, B, C, D (6 rats in each group). The rats were sacrificed at 8 h, 16 h, 24 h and 48 h, respectively. The changes of Brn3a expression in retina were observed by immunohistochemical method, and the changes of Brn3a mRNA in retina were observed by real time quantitative RT-PCR. Results: 1 Optical microscope: the retinal tissue structure of the experimental control eyes and normal eyes were clear, stained evenly, and the morphology of the cells was regular. Vacuolar degeneration occurred in some retinal ganglion cells (RGCs) at 8 hours after the model. After 16 hours, the number of (RGCs) in retinal ganglion cells decreased compared with the normal eyes, the arrangement was loose, some cells showed nuclear shrinkage, the degeneration of intracellular vacuoles became obvious, and the inner layer of retina became thinner. After 24 hours, the retinal ganglion cells decreased obviously, the nucleus became pyknosis, the degeneration of perinuclear vacuole was obvious, the arrangement of retinal nuclear layer cells was loose and disordered, and the thickness of retinal nuclear layer became thinner. 48 hours after the model, the retinal ganglion cells were few, occasionally visible, nuclear pyknosis, dense staining, vacuolar degeneration, irregular morphology, the structure of the retinal nuclear layer is more disorder, thickness is thinner. 2. Immunohistochemical staining: Brn3a was only expressed in the retinal ganglion cell layer. In experimental eyes, the expression of Brn3a was not significantly different from that of normal retina at 8 h after modeling. After 16 h of modeling, the expression of Brn3a RGCs had been reduced, and the number of Brn3a RGCs was significantly decreased after 24 h, and 48 h later. Brn3a RGCs distribution is sparse and sporadic. Compared with the control group, the expression of Brn3a RGCs in the experimental group was not significantly different from that in the blank group. 3. The expression of Brn3amRNA in the real-time quantitative PCR: group was significantly lower than that in the normal control group at 8 h after the establishment of the model (P0.05), which was 78.7% of the normal control group. After 16 hours, the expression level of 27.1g / 24 h in the normal control group was lower than that in the normal control group (11.5h / 48h), but the actual expression level was only 6.2% of the normal control group (P < 0.05). However, there was no significant difference in the expression of Brn3amRNA between the experimental control eyes and the blank control eyes (P0.05). Conclusion: NMDA can induce excitatory injury of rat retina, decrease the number of retinal ganglion cells and decrease the expression of Brn3a and mRNA in the retina.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R779.1
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