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目的 :观察大鼠视网膜缺血再灌注损伤 (retinalischemiareperfusioninjury ,RIR)中视网膜细胞的凋亡情况。方法 :利用前房高眼压灌注法 ,造成大鼠视网膜缺血 6 0分钟 ,然后恢复其供血 ,并分别于再灌注 12、2 4、4 8小时、7天后处死大鼠 ,摘取眼球作视网膜组织石蜡切片 ,用核苷酸末端转移酶介导的dUTP末端标记法 (terminaldeoxynucleotidyltransferasemediateddUTPnickend labeling ,TUNEL) ,亲和素 -生物素 -过氧化物酶技术 (avidinbiotinperoxidasecomplextechnique ,ABC)分别检测视网膜细胞的凋亡情况和P53 蛋白的表达和分布状况 ,同时对视网膜凋亡细胞进行计数并作统计学分析。结果 :RIR 12、2 4、4 8小时、7天后视网膜凋亡细胞数 (每高倍视野均数 )分别为 9 2± 0 912、2 5 6± 1 6 2 5、12 1± 1 6 0 9、0 0 2± 0 14 1。RIR 2 4小时时 ,P53 蛋白表达阳性。凋亡细胞及P53 阳性蛋白均出现在内核层 (innernuclearlayer ,INL)及神经节细胞层 (ganglioncelllayer ,GCL) ,而外核层 (outernuclearlayer,ONL)几乎无阳性表达。结论 :RIR中视网膜内层细胞存在着明显的凋亡征象。尤以RIR 2 4小时组最为显著。
Objective: To observe the apoptosis of retinal cells in retinal ischemia-reperfusion injury (RIR) in rats. Methods: Anterior chamber intraocular pressure perfusion was used to induce retinal ischemia in rats for 60 minutes, and then the blood supply to the retina was resumed. Rats were sacrificed 12, 24, 48, and 7 days after reperfusion, respectively. Retinal tissue paraffin sections were examined for retinal cell apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and avidin biotin peroxidase complex technology (ABC) And the expression and distribution of P53 protein. At the same time, the apoptosis of retinal cells were counted and analyzed statistically. Results: The number of retinal apoptotic cells (average number of high power field per horizons) at RIR 12,2 4,4 8 hours and 7 days was 9 2 ± 0 912, 25 6 ± 1 6 2 5, 12 1 ± 1 6 0 9 , 0 0 2 ± 0 14 1. P53 protein expression was positive at 4 hours of RIR2. Apoptotic cells and P53 positive proteins were found in the inner nuclear layer (INL) and ganglion cell layers (GCL), while the outer nuclear layer (outernuclear layer, ONL) almost no positive expression. Conclusion: There are obvious signs of apoptosis in the retinal inner layer cells in RIR. In particular, the RIR 2 4-hour group was the most prominent.