角膜穿通伤后视网膜钙蛋白酶、细胞骨架蛋白α-Ⅱ spectrin和NF200的分布及其相对含量  

Distribution and relative contents of m-calpain,cytoskeleton proteins α-Ⅱ spectrin and NF200 in retina after corneal penetrating injury

作　　者：黄燕[1] 吴滢[2] 宗海洋[2] 肖寿华[1] 张志坚[3] 

机构地区：[1]江苏大学附属医院眼科,江苏省镇江市212001 [2]江苏大学临床医学院,江苏省镇江市212003 [3]江苏大学基础医学与医学技术学院,江苏省镇江市212003

出　　处：《眼科新进展》2011年第10期922-925,929,共5页Recent Advances in Ophthalmology

摘　　要：目的观察角膜穿通伤后钙蛋白酶(m-calpain)、血影蛋白(α-Ⅱspectrin)及神经丝蛋白-200(neurofilament protein-200,NF200)在视网膜中的分布特征,检测骨架蛋白降解产物相对含量的动态变化,初步探讨角膜穿通伤后视网膜继发性损伤的病理机制。方法清洁级成年雌性SD大鼠50只,随机分为正常对照组和角膜穿通伤后6h、24h、48h、72h组,每组各10只。于眼球颞侧角膜缘用无菌注射器刺穿角膜制作角膜穿通伤模型。各组5只大鼠制作眼球切片,用相应抗体免疫荧光染色,观察m-calpain、α-Ⅱspectrin和NF200在视网膜中的分布特征;同时用免疫印迹法检测各组另外5只大鼠眼球壁组织中上述蛋白相对含量的动态变化。结果正常对照组m-calpain主要分布于视网膜节细胞,组织中蛋白相对含量较低;角膜穿通伤组组织中阳性细胞数量增多,蛋白相对含量亦增高,于24h达高峰,各组相比差异均有统计学意义(均为P<0.05)。正常对照组α-Ⅱspectrin和NF200主要分布于各种视细胞突起内,细胞界限和层次较清楚;角膜穿通伤组免疫荧光呈弥散分布,细胞界限和层次模糊不清;组织中α-Ⅱspectrin和NF200降解产物相对含量随m-calpain含量增高而增高,于24h达最大量,各组相比差异均有统计学意义(均为P<0.05)。结论角膜穿通伤可刺激视网膜细胞高表达m-calpain,后者对视网膜细胞骨架蛋白的降解是造成视网膜继发性损伤的重要因素。Objective To investigate the mechanisms of the second injury of retina after corneal penetrating injury by observing distribution of m-calpain,cytoskeleton proteins α-Ⅱ spectrin and neurofilament protein-200（NF200） and the dynamic changes of relative contents of degradation products of retinal nerve cell cytoskeleton proteins.Methods The 50 clear adult female rats were randomly divided into normal control group and corneal penetrating injury groups,which were subdivided into groups suffered from corneal penetrating injury for 6 hours,24 hours,48 hours and 72 hours,respectively.Corneal penetrating injury models were made with disposable sterilized syringe puncturing cornea at bitamporal corneal limbus.Five rats in each group were used to make eyeball slices.Corresponding antibody immunefluorescence staining was used to observe the distributions of m-calpain,α-Ⅱ spectrin and NF200 in retina.The relative contents of m-calpain,degradation products of α-Ⅱ spectrin and NF200 in the tissues of eye wall were detected by western blotting.Results The immunofluorescence staining of m-calpain in normal retina was mainly located in the retinal ganglion cells,and relative concentration of protein was low in tissues.The immunofluorescence was diffused in the retina of corneal penetrating injury groups.Western blotting results showed that the relative content of m-calpain was increased gradually and reached to its peak at 24 hours after corneal penetrating injury.There were statistical differences among groups（all P0.05）.The positive cells for α-Ⅱspectrin and NF200 were distinctly distributed in visual cell protuberant.Cell boundary and layers were clear.In corneal penetrating injury groups,immunofluorescence distributed diffusively,cell boundary and layers were not clear;Contents of α-Ⅱspectrin and NF200 increased with concentration of m-calpain,reached to peak at 24 hours,and there were statistical differences among groups（all P0.05）.Conclusions The corneal penetrating injury can activate the m-calpi

关 键 词：角膜穿通伤 钙蛋白酶 神经丝蛋白 血影蛋白 鼠 

分 类 号：R772.2[医药卫生—眼科]