眼缺血综合征大鼠视网膜凋亡相关蛋白的表达研究  

Expression of apoptosis-associated proteins in the rat retina of ocular ischemic syndrome

作　　者：谢国丽[1] 傅强[1] 赵露[1] 王艳玲[1] 

机构地区：[1]首都医科大学附属北京友谊医院眼科,100050

出　　处：《中华眼底病杂志》2015年第6期571-574,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（81173412）

摘　　要：目的观察眼缺血综合征（OIS）大鼠视网膜组织中促凋亡相关蛋白半胱氨酸蛋白酶（Caspase）-3、Caspase-8、B细胞淋巴瘤／白血病-2（Bcl-2）相关X蛋白（Bax），抑制凋亡相关蛋白Bcl-2的表达。方法Brown Norway大鼠30只随机分为实验组、对照组，每组均为15只大鼠。实验组大鼠行双侧颈总动脉结扎建立OIS动物模型；对照组大鼠游离双侧颈总动脉，但不结扎。建模后3个月，行苏木精-伊红染色，观察大鼠视网膜各层厚度及神经节细胞（RGC）数量；蛋白质免疫印迹法、实时定量聚合酶链反应检测，观察大鼠视网膜中Caspase-3、Caspase-8、Bax以及Bcl-2蛋白及基因表达。结果与对照组相比，实验组大鼠RGC数量明显减少，差异有统计学意义（t=3．059，P〈o．01）；视网膜总厚度（t=3．036）及内丛状层（t=3．715）、内核层（t=3．339）、外丛状层（t=3．341）厚度均明显降低，差异有统计学意义（P〈O．05）．外核层厚度无明显变化，差异无统计学意义（t=2．000，P〉0．05）。实验组大鼠视网膜Caspase-3、Caspase-8、Bax蛋白（F=17．036、7．787、11．431）及基因（F=11．162、17．763、12．183）表达均显著升高，差异有统计学意义（P〈0．05）；Bcl-2蛋白（F=10．298）及基因（F=12．047）表达显著下降，差异有统计学意义（P〈0．05）。结论凋亡相关蛋白在OIS大鼠视网膜组织中显著表达；促凋亡相关蛋白表达升高，抑制凋亡蛋白表达下降。Objective To observe the expression of Caspase-3, Caspase-8, Bcl-2, Bax in the rat retina of ocular ischemic syndrome （OIS）. Methods 30 Brown Norway rats were randomly divided into experimental group and control group, 15 rats in each group. The rats in experimental group were established a model through ligating the bilateral common carotid artery. At 3 months after modeling, the retinal thickness and ganglion cell （RGC） density were measured by hematoxylin eosin staining; the expression of Caspase 3, Caspase 8, Bax and Bcl-2 in the retina was measured by quantitative real-time reverse transcription polymerase chain reaction. Results The RGC density in control group and experimental group was 61.97 ± 9.07 and 38.1 ± 5.98, respectively. Compared to the control group, the RGC density was diminished in the experimental group （t=3. 059, P〈0.01）. A significant decrease was found in the total retina （t = 3. 036）, inner plexiform （t = 3. 715）, inner nuclear （t = 3. 339） and outer plexiform （t=3. 341） thickness （P〈0.05）. However, no change of the thickness was evident in the outer nuclear layers （t=2. 000, P〉0.05）. The levels of protein and RNA expression of Caspase 3, Caspase 8 and Bax in the retina were increased in experimental group （F=17. 036, 7. 787, 11. 431, 11. 162, 17. 763, 12. 183; P〈0.05）, while the Bcl 2 expression were decreased （F= 10.298, 12. 047; P〈0.05）. Conclusions There is obvious expression of apoptosis-associated proteins in the rat retina of OIS. Caspase 3, Caspase 8 and Bax expression are increased, while Bcl-2 expression are decreased.

关 键 词：缺血/病理生理学 细胞凋亡 半胱氨酸天冬氨酸蛋白酶 BCL-2相关X蛋白质 动物实验 

分 类 号：R771[医药卫生—眼科]