实验性自身免疫性葡萄膜炎眼球中自然杀伤细胞的致炎作用及其机制  

Inflammation-causing effects and mechanism of natural killer cells in experimental autoimmune uveitis rats

作　　者：李墨涵 鲍宁[1] 刘东伟[1] 陶黎明[1] 蒋正轩[1] 

机构地区：[1]安徽医科大学第二附属医院眼科,合肥230601

出　　处：《中华实验眼科杂志》2017年第9期799-804,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81300755）

摘　　要：背景实验性自身免疫性葡萄膜炎（EAU）是葡萄膜炎常见的动物模型，自然杀伤（NK）细胞是一种强烈的致炎细胞，但其在EAU中的作用及其机制仍有待研究。目的探讨EAU模型大鼠不同发病阶段视网膜组织中NK细胞的定位和分布及其机制。方法将36只SPF级Lewis大鼠应用随机数字表法随机分成对照组和造模后第6、9、12、16、21天组，每组各6只。造模后第6、9、12、16、21天组大鼠采用光感受器间维生素A类结合蛋白（IRBP）联合5mg／ml结核杆菌和完全福氏佐剂（CFA）乳化液双后足垫皮下注射，然后腹腔内注射400ng百日咳毒素免疫大鼠建立EAU大鼠模型。对照组大鼠双后足垫皮下注射生理盐水与等容量CFA乳化液，然后腹腔内注射400ng百日咳毒素。造模后每天用裂隙灯显微镜观察大鼠眼前段炎症反应过程，根据Caspi方法进行眼部炎症症状评分。分别于造模后第6、9、12、16、21天摘取各组大鼠眼球，采用苏木精一伊红染色法检测各组大鼠视网膜炎症反应和组织结构形态学改变；采用免疫荧光双标法检测大鼠视网膜中NK细胞的分布及浸润情况。另取25只Lewis大鼠应用随机数字表法随机分为造模后第0、3、6、9和12天组，每组5只大鼠，电动匀浆机破裂组织并匀浆为眼内液，采用实时荧光定量PCR法检测大鼠眼内液中NK细胞趋化因子CXCL10 mRNA和CXCL12 mRNA的表达。结果对照组大鼠眼前节未发现炎症反应。造模后第6天组大鼠虹膜血管扩张，随着造模后时间延长虹膜血管扩张明显，前房逐渐出现渗出或积脓，造模后第12天炎症反应达峰。视网膜组织病理学检查显示，对照组大鼠视网膜组织结构排列整齐，各EAU模型组大鼠造模后随时间延长均出现不同程度的视网膜结构排列紊乱，外核层细胞分离，层间组织松散，视细胞水肿且有炎性细胞浸润，以造模后第12天组最为严重。免�Background Experimental autoimmune uveitis （EAU） is a common animal model of uveitis. Natural killer （NK） cells have been confirmed to be a type of strong inflammation-causing cells, but its role in EAU is still studing. Objective This study was designed to explore the role and mechanism of NK cells in the pathogenesis of EAU. Methods Thirty-six SPF Lewis rats were randomly divided into expeimental control group and EAU 6-,9-, 12-, 16-, and 21-day groups （6 rats for each group）. Rats in EAU group received subcutaneous injection interphotoreeeptor retinoid binding protein （IRBP） combining 5 mg/ml tubercle bacillus with complete Freund＇s adjuvant （CFA） emulsion in foot pads, and then 400 ng pertussis toxin was intraperitoneally injeeted to extablish EAU models in the EAU 6-,9-, 12-, 16-, and 21-day group, and normal saline solution combined with CFA and 4-00 ng pertussis toxin was used in the same way in the experimental control group. The inflammatory response was observed by slit lamp daily after modeling and scored based on Caspi criteria. The eyeballs were extracted in 6,9,12, 16 and 21 days after modeling for retinal histopathological examination. Immunofluorescent double-staining was employed to detect and locate the expression of NK cells in the retina. In addition,25 model rats were divided into EAU 0-,3-, 6-, 9- and 12-day groups, with 5 rats for each group, and eyeballs were extracted to prepare tissue homogenate. The expression of CXCL10 mRNA, and CXCL12 mRNA NK cell chemokines, in the tissue homogenate was assayed by real-time quantitative PCR. The use and care of the rats followed Regulations for the Administration of Affair Concerning Experimental Animal by State Science and Technology Commission. Results No inflammatory sign in ocular anterior segment of the rats was seen in the experimental control group. The expansion of rat iris vessels was found in the EAU 6-day group, and exudes and hypopyon of the anterior chamber occurred in the EAU 9-day group and the inflammation peaked

关 键 词：自然杀伤细胞/免疫 葡萄膜炎/免疫 视黄醇结合蛋白类/免疫 自身免疫性疾病/病理 炎症 趋化因子 动物模型 大鼠 

分 类 号：R773.9[医药卫生—眼科]