新型自由基清除剂对鼠视神经挫伤后视网膜细胞的保护作用  

Protective effects of a new ROS scavenger on the retinal cells after optical nerve crash in rats

作　　者：周海燕[1] 高晨[2] 张景科[3] 周丽雅[4] 王小梅[5] 田蔓男[1] 魏志诚[1] 

机构地区：[1]兰州大学第一医院眼科 [2]兰州军区总医院安宁分院创伤外科,甘肃兰州730070 [3]兰州大学医学实验动物中心,甘肃兰州730000 [4]甘肃省人民医院神经内科电生理检查室,甘肃兰州730000 [5]兰州大学第一医院病理科,甘肃兰州730000

出　　处：《西安交通大学学报（医学版）》2014年第1期40-44,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：甘肃省科技厅技术研究与开发项目(No.1105TCYA004)~~

摘　　要：目的探讨大鼠视神经钳夹伤致视网膜细胞凋亡过程中活性氧(reactive oxygen speciesin,ROS)的含量和依达拉奉(edaravone,MCI-186)对视网膜细胞的保护作用。方法 SD大鼠192只,雌雄各半,随机分为正常组、假手术组、对照组、治疗组,每组各48只。对照组、治疗组用视神经钳夹法制作大鼠视神经夹挫伤模型;治疗组造模后腹腔注射依达拉奉30mg/kg,用生理盐水稀释后每隔12h腹腔注射给药。给药时间为30min,共14d。于实验的3、6、10、14d处死大鼠。2′,7′-二氯双氢荧光素双乙酸酯(DCFH-DA)为标记探针,通过流式细胞术定量检测各组不同时间点视网膜细胞内2′,7′-二氯双氢荧光素(DCF)的荧光强度;以AnnexinV/PI双染色法通过流式细胞术定量检测各组不同时间点视网膜细胞凋亡率;激光共聚焦显微镜观察细胞凋亡的形态特征;透射电镜观察各组鼠视网膜神经节细胞(retinal ganglion cells,RGCs)的超微结构改变。结果造模后3～14d,视网膜细胞中活性氧的表达量和细胞总凋亡率在3、6、10、14d时每个时间点对照组较治疗组和正常组明显升高,差异有统计学意义(P<0.05),假手术与正常组间相比均无显著性差异。透射电镜检查显示:治疗组大鼠RGCs细胞核、染色质、细胞器损伤较对照组有明显减轻。结论依达拉奉通过清除ROS,阻止了RGCs凋亡,具有一定的视网膜细胞保护作用。Objective To investigate the content of reactive oxygen species （ROS） during retinal cells apoptosis after optical nerve crash in rats and edaravone＇s protective effects on retinal cells. Methods A total of 192 Sprague-Dawley rats （96 males and 96 females） were randomly divided into normal group, sham group, control group and treatment group, with 48 rats in each group. In the control and the treatment group, the model of SD rat optic nerve crush was made with optic nerve crush method. In the treatment group, edaravone （30 mg/kg） diluted with normal saline was injected intraperitoneally after modeling every 12 hours with delivery time of 30 minutes for 14 days. The rats were killed at day 3, 6, 10 and 14 of the experiment. 2＇, 7＇-dichloro fluorescein pair of dihydroartemisinin acetate ester （DCFH-DA） was used as the labeled probe; fluorescence intensity of 2＇,7＇-dichloro fluorescein dihydrotestosterone （DCF） in retinal cells was checked by flow cytometry at different time points in the 4 groups. Apoptosis rate of the retinal cells was checked by double staining of AnnexinV/PI via flow cytometry at different time points in the 4 groups. Morphology of cell apoptosis was observed with laser confocal microscope. The ultrastructural changes of retinal ganglion cells （RGCs） were detected by TEM. Results At 3 d, 6 d, 10 d and 14 d, the expression of ROS and apoptosis rate of retinal cells in the control group were significantly increased at each time point compared with those in the treatment group and the normal group （P〈0.05）. There were no significant differences in all indexes between sham group and the normal group. Cellular damage in the treatment group abated obviously compared with that in the control group. Conclusion Edaravone prevents the apoptosis of retinal cells by eliminating ROS and thus has a protective effect on retinal cells.

关 键 词：自由基清除剂 视神经钳夹伤 视网膜细胞 活性氧 凋亡 超微结构 依达拉奉 

分 类 号：R774[医药卫生—眼科]