视网膜抗原免疫联合内毒素诱导的新的EAU小鼠模型  

Establishment of a new EAU animal model by endotoxin injection following immunization with IRBP emulsified in complete Freund adjuvant

作　　者：陈杰[1] 孙卫民[2] 王金泉[3] 宋斗[1] 张超雄[4] 徐晓[3] 

机构地区：[1]解放军第一一七医院眼科,杭州310013 [2]上海第二军医大学免疫学研究所,200433 [3]解放军第一一七医院病理科,杭州310013 [4]成都军区疾病预防控制中心,610021

出　　处：《中华实验眼科杂志》2014年第6期512-517,共6页Chinese Journal Of Experimental Ophthalmology

摘　　要：背景 葡萄膜炎的发病机制和治疗仍是目前的研究热点,但多年来该领域的基础研究仍是沿用传统的造模方法制备相关的动物模型,与人类的葡萄膜炎自然病程有较大偏差. 目的 本研究用大肠杆菌内毒素,即脂多糖（LPS）替代百日咳毒素（PTX）作为主要诱发因素,建立更符合人类自然发病环境的新型实验性自身免疫性葡萄膜视网膜炎（EAU）的动物模型,并与传统的造模方法进行比较,为研究该病的发病机制和有效的治疗方案提供实验依据.方法 6～8周龄的无特定病原体级雌性C57BL/6（H-2b）小鼠20只,按随机数字表法分为正常对照组、单纯内毒素注射组（EIU组）、多肽＋完全弗氏佐剂（CFA）注射组（EAU组）、多肽＋CFA＋LPS组（LPS-EAU组）.LPS-EAU组先用人类光感受器间维生素A类结合蛋白（IRBP 1-20）＋CFA免疫小鼠,免疫后第7天小鼠足底注射LPS,诱发小鼠EAU模型.采用组织病理学损害、眼球组织病理学评分、迟发型过敏反应、特异性淋巴细胞增生反应等评价指标对动物模型进行鉴定,并与LPS诱导的EIU及IRBP 1-20＋ CFA免疫诱导的EAU进行比较.结果 正常对照组小鼠虹膜睫状体及视网膜组织结构未见异常;EIU组小鼠虹膜睫状体可见轻微血管扩张、蛋白及纤维素渗出,但玻璃体和视网膜组织内未见血管异常及炎症反应;EAU组小鼠虹膜睫状体未见血管扩张及炎性渗出,但可见视网膜轻微血管周围炎及神经纤维层肿胀;LPS-EAU组小鼠视网膜结构紊乱,可见较多的炎性细胞浸润、光感受器细胞损伤及视网膜全层破坏.正常对照组小鼠和EIU组小鼠病理评分均为0分,EAU组病理评分为0.5分,而LPS-EAU组小鼠病理评分为3.0分,显著高于EAU组,差异有统计学意义（U=16.246,P=0.001）.LPS-EAU组小鼠耳廓增厚值为（35.60±0.55） tm,显著高于EIU组小鼠的（12.60±0.55）μm,差异均有统计学意义（q=23.003,P＜0.01）;但与EAU�Background The pathogenesis and management of human autoimmunity uveoretinitis is a focus in ophthalmology.For decades,a traditional experimental autoimmunity uveoretinitis （EAU） induced by pertussis toxin （PTX） was used for the basic investigation,which was thought to have a large deviation from the natural course of human autoimmunity uveoretinitis.Objective This study was to establish a new mice model of autoimmunity uveoretinitis which mimics the human autoimmunity uveoretinitis pathogenesis and offer a basis for the investigation and treatment of uveoretinitis.Methods Twenty 6-8 weeks old specific pathogen-free female C57BL/6 （H-2b） mice were randomized into normal control group,only endotoxin （lipopolysaccharide,LPS） induced uveitis group （endotoxin induced uveitis [EIU] group）,interphotoreceptor retinoid-binding protein （IRBP1-20） ＋complete Freund adjuvant （CFA） induced uveoretinitis group （EAU group） and IRBP＋CFA＋LPS induced uveoretinitis group （LPS-EAU group）.The mice of the EAU were only immunized with IRBP emulsified in CFA,and LPS-EAU group firstly were immunized with IRBP emulsified in CFA and then LPS was injected in the footpad of the mice on 7 days following immunization.The ocular pathological examination,histopathological scoring,delayed-type hypersensitivity and specific lymphocyte proliferating response were evaluated and compared with the EIU models,traditional EAU models without PTX and LPS-EAU models.The use and care of experimental animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results No inflammatory response was found in the iris,cilliary body and retina of mice in the normal control group.However,mild blood vessels dilation and fibrin exudation were seen in the iris and cilliary body of mice in the EIU group.In the EAU group,mild vasculitis and swelling of nerve fiber layer were exhibited in the retinas; while in the LPS-EAU group,severe disorder of ret

关 键 词：实验性自身免疫性葡萄膜视网膜炎 内毒素 脂多糖 动物模型 C57BL 6(H-2b)小鼠 

分 类 号：R774.1[医药卫生—眼科]