自身免疫性视网膜病变小鼠模型的建立及评价  

Establishment and evaluation of a murine model of autoimmune retinopathy

作　　者：刘谦[1] 周健[1] 武珅[1] 张子俊 张敬学[1] 曾惠阳[1] Liu Qian;Zhou Jian;Wu Shen;Zhang Zijun;Zhang Jingxue;Zeng Huiyang(Beijing Institute of Ophthalmology,Beijing Tongren Eye Center,Beijing Tongren Hospital,Capital Medical University,Beijing Key Laboratory of Ophthalmology and Visual Sciences,Beijing 100730,China)

机构地区：[1]首都医科大学附属北京同仁医院、北京同仁眼科中心、北京市眼科研究所、眼科学与视觉科学北京市重点实验室,100730

出　　处：《眼科》2023年第2期142-147,共6页Ophthalmology in China

基　　金：北京市自然科学基金(7192034);北京市眼科研究所突破计划项目(2019-2020);国家自然科学基金(81100675)。

摘　　要：目的诱导自身免疫性视网膜病变(AIR)小鼠模型,对其发生过程、病理及功能特征进行评价。设计实验研究。研究对象18只7~9周鼠龄C57BL/6J小鼠用于诱导AIR发生;同龄同种小鼠6只设立为对照组。方法用完全弗氏佐剂(CFA)乳化的小鼠重组恢复蛋白(recoverin)(CFA-recoverin)免疫小鼠作为诱导组,用CFA-PBS注射小鼠作为对照组。在免疫后的第0天和第2天注射百日咳毒素(PTX)破坏血-视网膜屏障。利用蛋白印迹法分析、多模态影像检测及组织病理学方法评价造模指标。主要指标诱导后3、6、8周小鼠血清recoverin抗体的表达、裂隙灯检查、彩色眼底照相、OCT、FFA或视网膜电图(ERG)的表现及视网膜组织学染色特征。结果诱导组小鼠在第3周出现血清recoverin抗体阳性;第6周抗体表达明显增加并开始出现双眼少量视网膜黄白色病灶,OCT示外层视网膜轻度受损;第8周视网膜浸润灶明显扩大,OCT显示外层视网膜连续性明显破坏。第8周FFA显示病灶区视网膜下明显荧光渗漏;ERG示视杆及视锥反应波振幅显著降低。第6周及第8周病理学显示不同程度外层视网膜结构破坏及炎性细胞浸润。结论重组recoverin蛋白皮下注射可成功诱导小鼠AIR发生,其眼部表现及病理学特征与AIR患者大致相似,是进行AIR研究的良好工具。Objective To establish and evaluate a murine model of autoimmune retinopathy.Design Animal experiment.Participants Eighteen C57BL/6J mice of 7~9 weeks old as experimental group and another six age-matched C57BL/6J mice as control group.Methods Mice in experimental group were immunized with recombinant mouse recoverin(200μg/mouse)in complete Freund’s adjuvant(CFA),while mice in control group were immunized with CFA-PBS.All of them were injected with 200 ng of pertussis toxin(PTX)day 0 and day 2 after immunization to facilitate cellular infiltration of the retina.The main indexes of the model were evaluated by western blotting,multi-modality imaging and pathological methods.Main Outcome Measures Presence of serum recoverin antibody,retinal findings and pathology were shown by slit-lamp examination,fundus photography,OCT,FFA,ERG,as well as HE staining of retinal cryosection at weeks 3,6 and 8 after immunization.Results Presence of yellow fleck lesions in the deep retina was observed from 6 weeks and significantly progressed at weeks 8 post-immunization of recoverin.Damage of outer retinal elements on OCT,reduced amplitude of ERG response and leakage of fluorescent dyes were shown at weeks 6 or/and 8 post-immunization.Intrusion of inflammatory cells into retinal tissue with various degree were also found on the cryo-section of weeks 6 and 8 AIR model.The AIR model was tested positive for serum recoverin antibody from weeks 3 post-immunization and keep high level at weeks 6 and 8 after immunization.Conclusions A murine AIR model immunized with recoverin protein was successfully established and evaluated in terms of retinal features,pathological changes and presence of serum recoverin antibody.Its similarity with AIR patients made it a good tool for investigating the pathogenesis of the disease.

关 键 词：自身免疫性视网膜病变 小鼠疾病模型 抗恢复蛋白抗体 抗视网膜抗体 

分 类 号：R774.1[医药卫生—眼科] R-332[医药卫生—临床医学]