重组人促红细胞生成素对小鼠视网膜光损伤的防护作用  

Edrythropoietin administration protects retina against light-induced retinal degeneration

作　　者：王红云[1] 牛膺筠[1] 王培嵩[1] 周占宇[1] 刘夫玲[1] 杨文毅[1] 张跃红[1] 

机构地区：[1]青岛大学医学院附属医院眼科,266003

出　　处：《中华眼科杂志》2005年第5期434-438,共5页Chinese Journal of Ophthalmology

摘　　要：目的　探讨重组人促红细胞生成素(rHEPO)通过小鼠血视网膜屏障的情况及其对视网膜光损伤的保护作用。方法　24只BALB/c小鼠腹腔注射rHEPO,采用酶联免疫吸附测定(ELISA)法检测小鼠视网膜中促红细胞生成素(EPO)的含量; 24只BALB/c小鼠建立光损伤动物模型,通过光镜和核苷酸末端转移酶介导的dUTP缺口标记(TUNEL)法观察实验组小鼠(12只,腹腔注射rHEPO)和对照组小鼠( 12只,腹腔注射生理盐水)视网膜细胞的变化情况。结果　腹腔注射rHEPO后2、4、6及8h小鼠100μg视网膜蛋白中EPO的含量分别为(0 .68±0 .24)、(1 .87±0 .37)、(0. 96±0 .24)及(0. 47±0. 13)mU,差异有统计学意义(F=2 .113,P<0 .05),在腹腔注射药物后4h视网膜中EPO的浓度达到高峰。随光照时间延长,光镜下可见对照组视网膜杆细胞的内、外节破坏明显加重,外核层逐渐变薄,出现核固缩,甚至核碎裂;实验组各观察时间点损伤变化均较轻,仅见感光细胞内、外节排列紊乱,形成空泡,外核层细胞排列稍紊乱,厚度无明显变化。荧光显微镜下观察,对照组视网膜外核层的凋亡细胞随光照时间延长不断增多,至光照后7d凋亡细胞数量减少;实验组视网膜外核层仅见少量凋亡细胞,光照后72h凋亡细胞的数量明显减少,至光照后7d几乎无凋亡细胞。实验组视网膜外核层凋亡细胞的数量?Objective To study whether recombinant human erythropoietin can pass through mice blood-retina barrier and the protective role in light-induced damage in retina. Methods After the injection of rHEPO, the content of rHEPO in 24 BALB/c mice retina was examined by enzyme linked immunosorbent assay (ELISA). 24 BALB/c mice were used to establish a light-induced damaged model, the difference of retina in rHEPO group and control group was compared using light microscope and TdT-mediated dUTP nick end labeling (TUNEL). Results The amount of retinal rHEPO in four deferent time points was (0.68±0.24)mU,(1.87±0.37)mU,(0.96±0.24) mU,(0.47±0.13) mU in 100 μg retinal total protein respectively by ELISA assay, there were statistical significances among groups. The density of rHEPO in the retina reached its peak at 4th hour after injection. Histology analysis: rHEPO group, at the 12th hour after light exposure the inner segment became condensed and disorganized. At the 36th hour the retina disorganized and vesiculated were seen in outer segments. At the 72th hour the inner and outer segments were damaged more seriously and the outer nuclear layer became thinner and denser. On the 7th day, the retinal outer nuclear layer became thinner and condenses. rHEPO group showed a minimal damage in every time points but outer nuclear layer disorganized and vesiculated in inner and outer segments. No obvious changes in retinal thickness. The apoptotic cells were detected by TUNEL. At the 12th hour after light exposure, there were the apoptotic cells in outer nuclear layer near outer plexiform layer. At 36th hour the numbers of apoptotic cells were increased, however at the 72th it was decreased obviously, only a few scattering apoptotic cells were revealed in the outer nuclear layer. Numbers of apoptotic cells between the rHEPO group and control group in outer nuclear layer were statistical significance ( P <0.01). Conclusions rHEPO can pass through the mice blood-retina barrier and rHEPO has neuroprotective effect on mice

关 键 词：重组人促红细胞生成素 视网膜光损伤 酶联免疫吸附测定(ELISA) 防护作用 促红细胞生成素(EPO) BALB/c小鼠 RHEPO 血视网膜屏障 凋亡细胞 腹腔注射药物 显微镜下观察 保护作用 时间延长 视网膜细胞 外核层 实验组 对照组 

分 类 号：R774.1[医药卫生—眼科]