NMDA所致大鼠視網(wǎng)膜興奮性損傷后Brn-3a表達(dá)的變化
發(fā)布時(shí)間：2018-12-13 19:46
【摘要】：目的：研究NMDA(N-methyl-D-aspartate, N-甲基-D-天門冬氨酸)所致大鼠視網(wǎng)膜興奮性損傷后Brn3a表達(dá)的變化。 方法：健康成年清潔級(jí)SD大鼠30只,隨機(jī)分為正常對(duì)照組(6只),實(shí)驗(yàn)組(24只)。實(shí)驗(yàn)組所有大鼠右眼玻璃體腔注射NMDA作為實(shí)驗(yàn)眼,左眼玻璃體腔注射PBS作為實(shí)驗(yàn)對(duì)照眼,隨機(jī)分成A, B, C, D四個(gè)組(每組6只)分別在造模后8h,16h,24h,48h處死,免疫組織化學(xué)法觀察視網(wǎng)膜Brn3a表達(dá)的變化,實(shí)時(shí)定量RT-PCR法觀察視網(wǎng)膜Brn3a mRNA的變化。 結(jié)果：1光學(xué)顯微鏡：實(shí)驗(yàn)對(duì)照眼和正常眼視網(wǎng)膜組織結(jié)構(gòu)層次清楚,染色均勻,細(xì)胞形態(tài)規(guī)整。實(shí)驗(yàn)眼在造模后8h,部分視網(wǎng)膜神經(jīng)節(jié)細(xì)胞(RGCs)內(nèi)出現(xiàn)空泡變性；造模后16h,視網(wǎng)膜神經(jīng)節(jié)細(xì)胞(RGCs)數(shù)目較正常眼減少,排列疏松,部分細(xì)胞出現(xiàn)核固縮,胞內(nèi)空泡變性逐漸明顯,視網(wǎng)膜內(nèi)核層變��；造模后24h,視網(wǎng)膜神經(jīng)節(jié)細(xì)胞明顯減少,核固縮,核周空泡樣變性明顯,視網(wǎng)膜內(nèi)核層細(xì)胞排列疏松紊亂,厚度變薄；造模后48h,視網(wǎng)膜神經(jīng)節(jié)細(xì)胞已較少,偶爾可見,核固縮,濃染,胞內(nèi)空泡樣變性,細(xì)胞形態(tài)不規(guī)則,視網(wǎng)膜內(nèi)核層結(jié)構(gòu)更加紊亂,厚度更薄。2、免疫組織化學(xué)：Brn3a只在視網(wǎng)膜神經(jīng)節(jié)細(xì)胞層中表達(dá),在實(shí)驗(yàn)眼,Brn3a在造模后8h表達(dá)與正常視網(wǎng)膜相比變化不明顯,造模16h后,Brn3a+RGCs已有減少,24h后,Brn3a+RGCs數(shù)目明顯減少,48h后,Brn3a+RGCs分布稀疏,零星可見。實(shí)驗(yàn)對(duì)照組Brn3a+RGCs較空白組Brn3a+RGCs表達(dá)無(wú)明顯差別。3、實(shí)時(shí)定量PCR：實(shí)驗(yàn)組Brn3amRNA表達(dá)在造模后8h較正常對(duì)照組減少(P0.05),為正常對(duì)照組的78.7%。在16h后明顯減少,為正常對(duì)照組的27.1%,24h后表達(dá)量為正常對(duì)照組的11.5%,48h表達(dá)繼續(xù)降低,實(shí)際表達(dá)量?jī)H為正常對(duì)照組的6.2%。而實(shí)驗(yàn)對(duì)照眼與空白對(duì)照眼視網(wǎng)膜中Brn3amRNA的表達(dá)量無(wú)明顯差別(P0.05)。 結(jié)論：NMDA可造成大鼠視網(wǎng)膜興奮性損傷,導(dǎo)致視網(wǎng)膜神經(jīng)節(jié)細(xì)胞數(shù)目減少,視網(wǎng)膜中Brn3a及其mRNA的表達(dá)減少。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R779.1
本文編號(hào)：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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R779.1
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