细胞间黏附分子-1在氪激光诱发的大鼠脉络膜新生血管中的动态表达变化  

Dynamic expression of intercellular adhesion molecule-I in laser-induced choroidal neovascularization in Brown Norway rats

作　　者：高小燕[1] 何守志[2] 

机构地区：[1]北方工业大学医院眼科,北京100144 [2]中国人民解放军总医院,全军眼科中心,北京100853

出　　处：《中华实验眼科杂志》2015年第12期1103-1107,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：全军“十五”科研基金资助项目（02M014）

摘　　要：背景细胞外基质（ECM）的种类繁多，具有重要的生物学功能，它在脉络膜新生血管（CNV）生长过程中的重要作用日益受到人们的关注，细胞问黏附分子-1（ICAM-1）就是其中之一。ICAM-1参与新生血管的生成，但目前CNV与ICAM-1相关性研究仍较少。目的观察ICAM-1在激光诱导的大鼠脉络膜新生血管（CNV）中的动态表达变化，探讨其在CNV形成过程中的作用，为CNV的靶向治疗提供依据。方法按照随机数字表法将48只健康雄性清洁级BN大鼠随机分为光凝后1、2、3、4、5、6、7和8周组，每组6只，每只大鼠任意选取一眼作为实验眼，另一眼作为正常对照。采用氪激光光凝法建立CNV大鼠模型，每组大鼠实验眼于光凝后相应时间点行荧光素眼底血管造影（FFA）检查，计算荧光素渗漏量（吸光度，A值），检查后各组摘取眼球制备球壁石蜡切片，采用苏木精-伊红染色法检测大鼠CNV面积；分别采用免疫组织化学法和原位杂交技术测定大鼠CNV中ICAM-1蛋白及其mRNA的表达，并计算其相对表达量（A值）。结果苏木精-伊红染色结果显示激光光凝后视网膜内外颗粒层连续性断裂并内陷，Bruch膜断裂，巨噬细胞迁移至光凝斑区，可见纤维母细胞及血管内皮细胞增生。免疫组织化学染色结果显示ICAM-1蛋白主要表达于血管内皮细胞、视网膜色素上皮（RPE）细胞和巨噬细胞中。原位杂交染色结果显示ICAM-1 mRNA主要表达于视网膜外颗粒层细胞中。视网膜光凝后不同时间组间CNV面积、荧光素渗漏量、ICAM-1蛋白及其mRNA相对表达量的总体比较差异均有统计学意义（F=194．43、178．84、739．08、2463．51，均P〈0．05）。随着光凝后时间的延长，大鼠CNV面积逐渐增加，荧光素渗漏量增多，ICAM-1蛋白及其mRNA相对表达量均呈逐渐增长的趋势，均于光凝后8周达到最高水平。结论大鼠CNV区ICAM-1的表达�Background Extracellular matrix （ECM） has various kinds of function. The important role of ECM during choroidal ncovascularization （ CNV ） types and important biological attention. Intercellular adhesion molecule-1 （ICAM-1） ,which is one of ECMs,involves in the formation of blood vessels,but the relations between CNV and ICAM-1 is still unknown. Objective This study was to observe the dynamic expression of ICAM-1 in krypton laser-induced CNV and explore the effect of ICAM-1 on CNV. Methods Forty-eight healthy male clean BN rats were randomly divided into post-photocoagulation 1-week group,2-week group, 3-week group,4- week group, 5-week group, 6-week group, 7-week group and 8-week group. Laser-induced CNV models were monocularly established and the fellow eyes served as the normal controls. Fundus fluorescein angiography （FFA） was carried out to quantitate the leakage degree （ absorbency）. The eyeballs were enucleated on various time points, and CNV areas were assessed by hematoxylin and eosin staining; immunohistochemistry and in situ hybridization assay were employed, respectively, for the detection of relative expression levels of ICAM-1 protein and mRNA （absorbency）. The use and care of the animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Committee. Results After laser photocoagulation,the retinal inner and outer nuclear layers were ruptured and invaginated. Disruption of Bruch membrane and macrophages migrating to the photocoagulation zone were seen under the optical microscope. The proliferation of fibroblasts and vascular endothelial cells was exhibited. ICAM-1 protein was mainly expressed in vascular endothelial cells,retinal pigment epithelium （RPE） cells and macrophages,and ICAM-1 mRNA was mainly expressed in the outer nuclear layer. Statistically significant differences were found in the fluorescence leakage degree, the relative expression levels of ICAM-1 protein and mRNA as well as CNV ar

关 键 词：脉络膜新生血管 细胞间黏附分子-1/代谢 免疫组织化学 原位杂交技术 疾病模型 BN大鼠 

分 类 号：R773.4[医药卫生—眼科]