谷氨酰胺对大鼠眼挫伤后视网膜细胞HSP27和NF-κB表达的影响

发布时间：2018-05-17 23:01

本文选题：谷氨酰胺 + 眼挫伤　；参考：《辽宁医学院》2012年硕士论文

【摘要】：目的探讨谷氨酰胺(Glutamine, Gln)对视网膜HSP27（Heat shock protein27）和NF-κB（Nuclear factor-kappa B）的调节作用，明确Gln对挫伤大鼠视网膜保护作用机制，以期为临床治疗视网膜挫伤提供实验依据。方法 SD雄性大鼠45只，5只用于正常对照组。余下40只用于制备眼挫伤模型。正常组不处理，模型组和Gln组仿Allen’s重击法制作视网膜挫伤模型致左眼挫伤。模型制备成功后，40只大鼠随机分为2组，视网膜挫伤模型组(模型组)和Gln处理组(Gln组)。后两组又分为12h，24h，3天和7天4个时间段，每组每个时间段大鼠5只。伤后第2天，Gln组腹腔注射Gln（0.5g/kg），每日一次，模型组用同体积的生理盐水腹腔注射。Gln组、模型组各时段于给药给水停止后第2天取材，所有组别的大鼠均取左眼。应用HE染色方法观察各组视网膜各层细胞的形态改变；应用免疫组织化学方法检测视网膜组织中HSP27和NF-κB蛋白的表达变化。结果 1、视网膜各层组织病理学改变 HE染色观察可见模型组与Gln组视网膜全层细胞数减少，视网膜厚度较正常组明显变薄，各层细胞排列紊乱，细胞水肿；正常对照组视网膜各层细胞数量多，排列整齐，形态较完整。 2、HSP27蛋白检测结果光镜下HSP27蛋白免疫反应产物呈棕黄色。HSP27蛋白在正常组视网膜神经节细胞层、内核层胞浆内有表达，在外核层和感光细胞细胞层的胞浆内表达极少。损伤12h后，在模型组和Gln干预组，HSP27蛋白在视网膜的表达变化不明显，与正常组比较，无统计学意义。损伤24h后，模型组HSP27蛋白的表达明显下降，明显低于正常组（P0.05）3d后，HSP27蛋白表达降至最低，7d后，其表达有所恢复，但仍高于正常组（P0.05）。在Gln组，HSP27蛋白表达变化趋势与模型组相似，但在24h、3d、7d, HSP27蛋白表达较模型组明显增强，其蛋白阳性产物的平均光密度值(Mean optical density, MOD)明显高于模型组（P0.01）。 3、NF-κB蛋白检测结果 NF-κB蛋白免疫反应产物呈棕黄色，在神经节细胞层、内核层和外核层及感光细胞层的胞浆均有表达。正常组大鼠视网膜组织中有适量NF-κB蛋白表达。在模型组，损伤后12h，NF-κB蛋白在视网膜的表达变化不明显，与正常组比较，无统计学意义。损伤后24h，模型组NF-κB蛋白的表达升高，明显高于正常组（P0.01）；损伤后3d，NF-κB蛋白表达达峰值，至7d后，其表达略有下降，但仍高于正常组（P0.05）。在Gln组，除12h外，NF-κB蛋白表达在24h、3d、7d均较模型组明显降低，其蛋白阳性产物的平均光密度值(Mean optical density,MOD)明显低于模型组（P0.01）。结论 HSP27和NF-κB可能参与了眼挫伤视网膜病变过程；谷氨酰胺上调了挫伤视网膜神经细胞HSP27蛋白表达，，下调了NF-κB蛋白表达，这可能是谷氨酰胺对眼挫伤视网膜病变产生保护作用的机理之一。
[Abstract]:Purpose To investigate the regulatory effects of glutamine Glutamine27 (Glutamine27) and NF- 魏 B(Nuclear factor-kappa B on retinal contusion in rats, and to clarify the protective mechanism of Gln on retina contusion in order to provide experimental evidence for clinical treatment of retinal contusion. Method 45 SD male rats were used as normal control group. The remaining 40 cases were used to establish the model of eye contusion. The model group and Gln group made retinal contusion model by Allen's method. After the establishment of the model, 40 rats were randomly divided into two groups: retinal contusion model group (model group) and Gln treatment group. The latter two groups were divided into two groups: 12 h, 24 h, 3 days and 7 days, with 5 rats in each group. On the second day after injury, the GLN group was injected intraperitoneally with Glnn 0.5g / kg / kg, once a day. The model group was injected intraperitoneally with the same volume of normal saline. The rats in the model group were collected on the second day after the drug supply stopped, and the left eyes were taken from all groups of rats. The morphological changes of retinal cells in each group were observed by HE staining and the expression of HSP27 and NF- 魏 B in retinal tissues were detected by immunohistochemical method. Result 1. Histopathological changes in all layers of retina He staining showed that the number of whole retinal cells in the model group and Gln group decreased, the thickness of the retina became thinner than that in the normal group, the cells in each layer were disordered and the cells were edema, while in the normal control group, the number of cells in each layer of the retina was more than that in the normal control group. The shape is relatively complete. Detection of HSP27 protein Under light microscope, the immunoreactive product of HSP27 protein was brownish yellow. HSP27 protein was expressed in the retinal ganglion cell layer of normal group, and in the cytoplasm of nuclear layer, but in the outer nuclear layer and photosensitive cell layer. After 12 hours of injury, the expression of HSP27 protein in the retina of the model group and Gln intervention group was not significantly changed, and there was no significant difference between the model group and the normal group. After 24 hours of injury, the expression of HSP27 protein in the model group was significantly lower than that in the normal group. The expression of HSP27 protein in the model group was significantly lower than that in the normal group after 3 days, and the expression of HSP27 protein in the model group was lower than that in the control group. After 7 days of injury, the expression of HSP27 protein recovered, but still higher than that in the normal group. The change trend of HSP27 protein expression in Gln group was similar to that in model group, but the expression of HSP27 protein was significantly higher than that in model group at 24 h or 3 d, and the mean optical density of the protein positive product was significantly higher than that in model group (P 0.01). 3detection of NF- 魏 B protein The immunoreactive products of NF- 魏 B protein were brown and expressed in the cytoplasm of ganglion cell layer, nuclear layer, outer nuclear layer and photosensitive cell layer. In normal group, NF- 魏 B protein was expressed in retina tissue. In the model group, the expression of NF- 魏 B protein in the retina was not significantly changed at 12 h after injury, but there was no statistical significance compared with the normal group. The expression of NF- 魏 B protein in the model group was significantly higher than that in the normal group at 24 h after injury, and reached a peak at 3 days after injury, and decreased slightly at 7 days after injury, but still higher than that in the normal group. In the Gln group, the expression of NF- 魏 B protein was significantly lower than that of the model group on the 3rd day after 24 h, and the mean optical density of the protein positive product was significantly lower than that of the model group (P 0.01). Conclusion HSP27 and NF- 魏 B may be involved in the process of eye contusion retinopathy, glutamine upregulated the expression of HSP27 protein and down-regulated the expression of NF- 魏 B protein in retinal nerve cells. This may be one of the mechanisms of the protective effect of glutamine on ocular contusion retinopathy.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R779.1

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