实验性自身免疫性葡萄膜炎小鼠视网膜蛋白差异表达及生物信息分析  

Differential expression and bioinformation analysis of retinal proteins in mice with experimental autoimmune uveitis

作　　者：陈爽 邵先锋 张智慧 陈努 吴凌子 崔雪雪 李筱荣 张晓敏 Chen Shuang;Shao Xianfeng;Zhang Zhihui;Chen Nu;Wu Lingzi;Cui Xuexue;Li Xiaorong;Zhang Xiaomin(Tianjin Key Laboratory of Retinal Functions and Diseases,Eye Institute and School of Optometry,Tianjin Medical University Eye Hospital,Tianjin 300384,China)

机构地区：[1]天津医科大学眼科医院眼视光学院眼科研究所,天津市视网膜功能与疾病重点实验室,300384

出　　处：《中华实验眼科杂志》2019年第12期949-955,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目(81671642)。

摘　　要：目的观察实验性自身免疫性葡萄膜炎(EAU)小鼠视网膜蛋白质表达情况并探讨自身免疫性葡萄膜炎发生的可能分子机制。方法取12只SPF级雌性C57BL/6J小鼠采用随机数字表法随机分为模型组和正常对照组,每组6只。模型组小鼠皮下注射人光感受器间维生素A类结合蛋白(IRBP)651-670建立EAU模型,通过直接检眼镜观察眼底,采用OCT仪和组织病理学染色检测视网膜改变。在免疫后第18天收集2个组小鼠视网膜,进行视网膜蛋白提取、蛋白酶切、质谱检测、数据解析和生物信息学分析。结果成功建立EAU小鼠模型,免疫后第10天模型鼠视网膜出现局灶性损伤,免疫后第18天视网膜炎症反应达峰值,视网膜出现水肿、脱离,玻璃体液中有大量炎性细胞渗透。蛋白质组学结果显示,共鉴定到蛋白质4458个,其中差异表达蛋白(变化倍数>1.5,P<0.05)有522个。在这些差异表达蛋白中,上调的蛋白有147个,包括Stat1、Stat3等;下调的蛋白有375个,包括Gucy2f、Rho等。经基因本体(GO)分析和Reactome通路分析,这些差异蛋白主要与血小板活化、整合素信号、T细胞活化、光传导级联、Toll样受体等密切相关。结论EAV与Stat1、Stat3等蛋白的异常表达,以及血小板活化、整合素信号等信号通路的异常调节相关。Objective To observe the expression of retinal proteins in experimental autoimmune uveitis(EAU)mice and to explore the possible molecular mechanism of autoimmune uveitis.Methods Twelve female C57BL/6J mice were randomly divided into model group and normal control group,6 mice in each group.In the model group,the EAU model was established by subcutaneous injection of human interphotoreceptor retinoid-binding protein(IRBP)651-670.The fundal change of EAV mice was assessed by direct ophthalmoscope,OCT and histopathological staining.At 18 days after immunization,the retinas of the two groups were taken for retinal protein extraction,protein restriction enzyme digestion,mass spectrometry detection,data analysis,and bioinformatics analysis.This study was approved by the experimental animal Ethics Committee of Tianjin Medical University Eye Hospital(TJYY2018070113).The feeding and use of experimental animals follow the ARVO statement.Results The EAU mouse model was successfully established.At 10 days after immunitation,the retina of EAV mouse was damaged.At 18 days after immunization,retinal edema and infiltration of inflammatory cells into vitreous were observed.Proteomic results showed that a total of 4458 proteins were identified in this study,of which 522 were differentially-expressed proteins(fold change>1.5,P<0.05).Among these differentially-expressed proteins,147 proteins including Stat1 and Stat3 were up-regulated and 375 proteins including Gucy2f and Rho were down-regulated.These differentially-expressed proteins were closely related to platelet activation,integrin signaling,T cell activation,the phototransduction cascade,Toll-like receptor and so on.Conclusions EAU is related to the abnormal expression of Stat1,Stat3 and other proteins,as well as the abnormal regulation of platelet activation and integrin signal pathway.

关 键 词：葡萄膜炎/免疫 视网膜 自身免疫性疾病 蛋白组学 C57BL近交系小鼠 

分 类 号：R73[医药卫生—肿瘤]