中文摘要：

目的研究内质网应激介导凋亡途径的标志物生长停滞及DNA损伤基因153（GADD153）在大鼠实验性视网膜脱离后不同时期的基因与蛋白表达水平并探讨内质网应激与视网膜脱离后细胞凋亡发生的关系。方法对照实验研究。Wistar大鼠88只（88只眼），采用计算机随机数字表法，分为正常对照组和实验组，实验组包括视网膜脱离后1／2、1、2、4、8、16及32d组；每组各11只鼠（11只眼），通过在视网膜下注射透明质酸钠的方法，建立视网膜脱离模型。在视网膜脱离后1／2、1、2、4、8、16及32d分别摘除眼球。应用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL），检测视网膜细胞凋亡情况；采用半定量逆转录聚合酶链反应（RT-PCR）法，检测GADD153 mRNA的表达水平；应用免疫印迹法，检测视网膜组织中GADD153蛋白表达水平；采用免疫荧光和激光共焦显微镜技术，观察GADD153蛋白在视网膜各层细胞的分布。应用SPSS10．0统计学软件进行数据分析。对各组鼠视网膜凋亡细胞百分比和GADD153 mRNA表达水平的比较，采用Kruskal Wallis检验；GADD153蛋白表达水平的比较，采用One-Way ANOVA检验，以P〈0．05作为差异有统计学意义。结果大鼠视网膜脱离后凋亡细胞主要集中在光感受器细胞层，凋亡高峰出现在视网膜脱离后2—4d，8d后显著减少，组间视网膜细胞凋亡百分比差异有统计学意义（x^2=22．423，P〈0．05）；视网膜脱离后1／2、1、2及4d，视网膜GADD153 mRNA表达水平显著升高（x^2=27．223，P〈0．05）；GADD153蛋白表达水平亦显著升高（F=16．052，P〈0．05），主要表达部位在光感受器细胞层。结论视网膜脱离后内质网应激标志物GADD153被激活，并在视网膜组织表达水平增高，其表达状态与视网膜细胞的凋亡时间和发生位置相一致；内质网应激介导的凋亡途径参与了视网膜脱离?

英文摘要：

Objective To determine whether the endoplasmic reticulum stress participates into the apoptosis process of the retina during experimental retinal detachment-by detecting the level of mRNA and protein of the GADD153, the marker of endoplasmic reticulum stress. Methods Completely random design is applied. Experimental retinal detachment was created in the left eyes of 77 wistar rats by injecting hyaluronic acid into the subretinal space. Rats were sacrificed at 1/2 d, 1 d, 2 d, 4 d, 8 d, 16 d and 32 d after creation of retinal detachment. Apoptosis of retinal cells was detected by TdT-mediated fluorescein-16- dUTP nick-end labeling （TUNEL） assay. GADD153 mRNA expression in the retina was determined using semi-quantitative reverse transcription polymerase chain reaction （RT-PCR）. The GADD153 protein expression was determined using western blotting. Retinal sections were studied by immunoflurescence labeling and confocal microscopy. The SPSS 10. 0 software was applied to analyze data; Kruskal wallis test was applied to analyze the data about the apoptotic rate of retinal cells and the expression of GADD153 mRNA; One-way ANOVA test was applied to analyze the data about expression of GADD153 protein. P 〈0. 05 represents statistically significant difference. Results TUNEL positive staining cells were mainly presented in photoreeeptor cell layer, peaked at 2-4 days and markedly decreased at 8 days after retinal detachment. The difference of apoptotie retinal cell rate between groups is significant （x^2 = 22. 423, P 〈 0. 05） ;The expression of retinal GADD153 mRNA was significantly increased at 1/2 d, 1 d, 2 d, 4 d after retinal detachment（ x^2 = 27. 223, P 〈 0. 05 ） ; The expression of retinal GADD153 protein was significantly increased at these days（ F = 16. 052, P 〈 0. 05 ）. GADD153-positive cells were located in the photoreeeptor cell layer. Conclusions GADD153, the maker of endoplasmic reticulum stress mediated apoptosis, is activated and over-expressed, associated with the occurrence of a