TNF-α在NMDA诱导大鼠视网膜兴奋性损伤中的表达

发布时间：2018-06-12 13:48

本文选题：N-甲基天冬氨酸 + 肿瘤坏死因子α　；参考：《青岛大学》2011年硕士论文

【摘要】：目的：研究NMDA诱导大鼠视网膜兴奋性损伤中TNF-α的表达规律。方法：健康成年清洁级Wistar大鼠42只,随机分为正常对照组(6只)、PBS组(6只)和NMDA组(30只)。NMDA组所有大鼠均随机取一只眼球玻璃体腔注射2μl80mmol/l NMDA制作视网膜兴奋性损伤模型。PBS组大鼠随机取一只眼玻璃体腔注射2μl PBS。正常对照组不做任何处理。NMDA组大鼠在玻璃体腔注射后2h,8h,16h,24h,48h处死。HE染色光镜下观察各组视网膜组织的形态学变化,免疫组织化学染色方法观察视网膜TNF-α表达的变化,实时定量RT-PCR方法观察视网膜TNF-αmRNA表达的变化。结果：1 HE染色：正常对照组和PBS组大鼠视网膜组织结构层次清楚,染色均匀,细胞形态规则。NMDA组造模后2h,神经纤维层和神经节细胞开始水肿；造模后8h,水肿最严重,视网膜增厚；造模后16h,水肿较前减轻,排列疏松,形态不规则,数量减少；造模后24h,神经节细胞数目减少,排列紊乱,神经纤维层、神经节细胞层变薄；造模48h,神经节细胞稀少,神经纤维层和神经节细胞层萎缩,视网膜变薄.2免疫组织化学染色：正常对照组及PBS组视网膜未见TNF-α阳性表达。NMDA组造模后2h,TNF-α的表达量迅速上调(P0.05),造模后8h,TNF-α表达量达高峰(P0.05),造模后16h表达量降低,但仍高于对照组(P0.05)。造模后24h,48h TNF-α表达量与16h无差别(p0.05),较正常对照组增多(P0.05)。TNF-α阳性表达主要位于神经纤维层、神经节细胞层及内丛状层。3实时定量RT-PCR:正常对照组和PBS组大鼠视网膜未见TNF-αmRNA表达。NMDA组造模后2h,TNF-αmRNA水平较正常对照组上调(P0.05),造模后8h, TNF-αmRNA水平达高峰(P0.05),造模后16h表达量降低,但仍高于对照组(P0.05)。造模后24h,48h TNF-αmRNA表达量与16h无差别(p0.05),较正常对照组增多(P0.05)。结论：TNF-α在NMDA诱导大鼠视网膜兴奋性损伤中表达增加,该因子在视网膜兴奋性损伤中可能起重要作用。
[Abstract]:Objective: To study the expression of TNF- alpha in NMDA induced retinal excitability injury in rats.
Methods: 42 healthy adult clean Wistar rats were randomly divided into normal control group (6 rats), group PBS (6) and group NMDA (30), all rats of group.NMDA were randomly selected to take one eye glass cavity injection 2 mu l80mmol/l NMDA to make retinal excitatory injury model.PBS group, and the rats were randomly selected to take one eye glass cavity and injected 2 mu L PBS. normal control group randomly. No treatment was done to observe the morphological changes of retina tissue in each group of.NMDA rats after the injection of 2h, 8h, 16h, 24h, 48h after the injection of the vitreous cavity. The changes in the expression of TNF- alpha in retina were observed by immunohistochemical staining, and the change of mRNA expression of retina TNF- alpha was observed in real-time quantitative RT-PCR method.
Results: 1 HE staining: the retinal tissue structure of the normal control group and the PBS group was clear, the staining was uniform, the cell morphology rule of the.NMDA group was 2h, the nerve fiber layer and the ganglion cells began to dropout; after the model, 8h, the edema was the most serious, the retina thickened; after the model, the edema was reduced, the arrangement was loose, irregular and quantity. 24h, the number of ganglion cells decreased, the number of ganglion cells decreased, the nerve fiber layer, the ganglion cell layer thinned, the model 48h, the ganglion cells were scarce, the nerve fiber layer and the ganglion cell layer atrophied, and the retina thinned.2 immunohistochemical staining: the normal control group and the PBS group did not see the TNF- alpha positive expression of the.NMDA group. 2H, TNF- alpha expression increased rapidly (P0.05), 8h, TNF- alpha expression reached peak (P0.05), and 16h expression decreased after modeling, but it was still higher than that of control group (P0.05). The expression of 24h, 48h TNF- alpha was not different from that of 16h (P0.05), and the positive expression of 48h TNF- was mainly in the nerve fiber layer, ganglion cell layer and within the normal control group. There was no 2h in the retina of the plexiform.3 RT-PCR: normal control group and the rat retina of the PBS group, and the mRNA level of the TNF- alpha mRNA was higher than that of the normal control group (P0.05). The TNF- alpha level reached the peak after the model, and the expression of TNF- alpha was decreased, but it was still higher than that of the control group. There was no difference between 16h and P0.05 (P0.05), which was more than that in normal control group (P0.05).
Conclusion: TNF- alpha is increased in NMDA induced rat retinal excitability injury, which may play an important role in retinal excitability injury.
【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】：R774.1

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