眼科 ›› 2023, Vol. 32 ›› Issue (2): 142-147.

• 论著 • 上一篇    下一篇

自身免疫性视网膜病变小鼠模型的建立及评价

刘谦  周健  武珅  张子俊  张敬学  曾惠阳    

  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
  • 收稿日期:2022-11-11 出版日期:2023-03-25 发布日期:2023-03-25
  • 通讯作者: 曾惠阳,Email: zhydr@hotmail.com
  • 基金资助:
    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)

Establishment and evaluation of a murine model of autoimmune retinopathy

Liu Qian, Zhou Jian, Wu Shen, Zhang Zijun, Zhang Jingxue, Zeng Huiyang   

  1. 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
  • Received:2022-11-11 Online:2023-03-25 Published:2023-03-25
  • Contact: Zeng Huiyang, Email: zhydr@hotmail.com
  • Supported by:
    Beijing Municipal Natural Science Foundation (7192034); Key Research Program of Beijing Institute of Ophthalmology (2019-2020); National Natural Science Foundation of China (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。(眼科,2023,32: 142-147)

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

Abstract: 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 (200 μg/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. (Ophthalmol CHN, 2023, 32: 142-147)

Key words: autoimmune retinopathy, murine model, recoverin antibody, anti-retinal antibody