实验性视网膜脱离后光感受器细胞的坏死性凋亡  

Photoreceptor necroptosis in experimental retinal detachment

作　　者：董凯[1] 周恩亮[1] 朱子诚[1] 文磊[1] 冯敬仰 闫泉[2] 柯根杰[1] 孙晓东[2] 

机构地区：[1]安徽医科大学附属省立医院眼科,合肥230001 [2]上海交通大学附属上海市第一人民医院眼科

出　　处：《中华眼底病杂志》2014年第4期378-380,共3页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（NSFC81170861）;安徽省自然科学基金（AHNSF1408085QH159）

摘　　要：目的 观察实验性视网膜脱离后光感受器细胞坏死性凋亡的形态学特点,初步探讨其发生机制.方法 雄性健康的Sprague-Dawley大鼠60只,右眼视网膜下注入1％透明质酸纳50μl建立视网膜脱离模型作为实验组,左眼不作处理为对照组.建模后3d,采用电子显微镜观察光感受器细胞的死亡方式及形态学特点.建模后3d,提取大鼠视网膜组织,采用蛋白质免疫印迹法检测裂解的半胱氨酸蛋白酶（Caspase）8的相对蛋白表达;采用免疫沉淀法检测磷酸化受体相互作用蛋白1（p-RIP1）的相对蛋白表达.结果 电子显微镜观察发现,视网膜脱离后光感受器细胞的死亡方式主要为凋亡和坏死.同时还可观察到光感受器细胞的坏死性凋亡,其形态学与坏死细胞类似.表现为细胞肿胀,质膜不完整、破裂,染色质浓集和边聚,且伴有明显的自噬泡和空泡.蛋白质免疫印迹法检测结果显示,实验组、对照组裂解的Caspase 8蛋白相对表达分别为0.78±0.03、0.06±0.01,实验组裂解的Caspase 8蛋白相对表达较对照组明显增加.两组裂解的Caspase 8蛋白相对表达比较,差异有统计学意义（F=4 023.21,P＜0.05）.免疫沉淀法检测结果显示,实验组、对照组p-RIP1蛋白相对表达分别为0.23±0.03、0.14±0.02,实验组p-RIP1蛋白相对表达较对照组明显增加.两组p-RIP1蛋白相对表达比较,差异有统计学意义（F=56.44,P＜0.05）.结论 实验性视网膜脱离后光感受器细胞坏死性凋亡表现为细胞肿胀,质膜不完整、破裂,染色质浓集和边聚,且伴有明显的自噬泡和空泡;其发生机制与p-RIP1表达增加有关.Objective To observe the morphological characteristics of photoreceptor necroptosis in experimental retinal detachment,and explore the mechanism.Methods A total of 60 Sprague-Dawley male rats were included in this study.Retinal detachment were induced in the right eyes with 1％ sodium hyaluronate （50 μl） injection （experimental group）,while the left eyes received no treatment （control group）.At 3 days after modeling,the morphological characteristics of photoreceptor cell were observed by electron microscopy.Cleaved Caspase 8 and phosphorylation receptor-interacting protein 1 （p-RIP1） were measured by Western blot and immunoprecipitation.Results At 3 days after modeling,photoreceptor necroptosis showed the following morphological features：chromatin condensation,severe vacuolation,early loss of plasma membrane integrity,and many autophagosomes.Western blot showed that the protein expression of cleaved Caspase 8 were 0.78 ± 0.03,0.06 ± 0.01 in experimental group and control group respectively,which was significantly different （F=4 023.21,P＜0.05）.Immunoprecipitation showed that the protein expression of p-RIP1 were 0.23±0.03,0.14±0.02 in experimental group and control group respectively,which was significantly different （F =56.44,P ＜ 0.05）.Conclusions Photoreceptor necroptosis showed chromatin condensation,severe vacuolation,early loss of plasma membrane integrity,and many autophagosomes.Necroptosis activation was associated with the increase of RIP1 phosphorylation.

关 键 词：视网膜脱离 病理生理学 光感受器细胞 细胞凋亡 动物实验 

分 类 号：R774.1[医药卫生—眼科]