ROS信号通路调控NLRP3炎症小体在自身免疫性葡萄膜炎发生中的作用  

Mechanism of ROS signaling pathway regulating NLRP3 inflammatory body in autoimmune uveitis

作　　者：王俞方 肖启国 王智 周俊 郑芳 WANG Yufang;XIAO Qiguo;WANG Zhi;ZHOU Jun;ZHENG Fang(Department of Ophthalmology,the Second Hospital of University of South China,Hengyang 421001,Hunan Province,China)

机构地区：[1]南华大学附属第二医院眼科,湖南省衡阳市421001

出　　处：《眼科新进展》2020年第6期527-532,共6页Recent Advances in Ophthalmology

基　　金：湖南省衡阳市科技局资助项目(编号S2018F9031022227)。

摘　　要：目的探讨活性氧簇(ROS)信号通路调控核苷酸结合寡聚化结构域样受体(nucleotide binding and oligomerization domain-like receptor,NLRP3)炎症小体的表达在自身免疫性葡萄膜炎(autoimmune uveitis,AU)发生中的作用。方法健康大鼠共40只,随机分为空白对照组、模型组、空载体转染组(空载体组)和NLRP3-shRNA沉默组(沉默组)各10只;沉默组构建慢病毒载体pLenti6.3-EmGFP-NLRP3-shRNA,空载体转染组注射等量空载体,感染时间72 h。结果ELISA结果发现,模型组和空载体组造模成功后10 d、13 d、16 d、19 d的NLRP3、IL^-1β和IL^-18水平显著高于沉默组,沉默组高于空白对照组(P<0.05)。免疫组织化学结果发现,模型组和空载体组12 d和20 d的NLRP3、凋亡相关斑点样蛋白(ASC)、pro-Caspase-1、IL^-1β和IL^-18阳性表达率显著高于沉默组,沉默组高于空白对照组(P<0.05)。PCR结果发现,模型组和空载体组造模成功后12 d和20 d的NLRP3 mRNA水平显著高于沉默组,沉默组高于空白对照组(P<0.05)。Western blot结果发现,模型组和空载体组造模成功后12 d和20 d的NLRP3、ASC、pro-Caspase-1、IL^-1β和IL^-18,硫氧还蛋白和硫氧还蛋白互作蛋白相对表达水平显著高于沉默组,沉默组高于空白对照组(P<0.05)。结论NLRP3炎症小体及下游炎症因子表达上调可能参与了AU的发生,可能介导ROS信号通路发挥作用。Objective To explore the effect of nucleotide binding oligomerization domain-like receptor(NLRP3)inflammatory body regulated by reactive oxygen species(ROS)signaling pathway in pathogenesis of autoimmune uveitis(AU).Methods A total of 40 healthy rats were divided randomly into blank control group,model group,blank vector transfected group(blank vector group)and NLRP3-shRNA silencing group(silencing group),10 rats in each group.The silencing group constructed lentiviral vector pLenti6.3-EmGFP-NLRP3-shRNA,the blank vector group injected the same amount of blank vector for 72 h.Results ELISA result showed that levels of NLRP3,IL^-1βand IL^-18 in model group and blank vector group were significantly higher than those in silencing group on 10th,13th,16th and 19th days,and were the lowest in blank control group(P<0.05).Immunohistochemical results showed that positive expression rates of NLRP3,apoptosis-related spot-like protein(ASC),pro-Caspase-1,IL^-1βand IL^-18 in model group and blank vector group on 12th and 20th days were significantly higher than those in silencing group,and were the lowest in blank control group(all P<0.05).PCR results showed that levels of NLRP3 mRNA in model group and blank vector group on 12th and 20th days were significantly higher than those in silencing group,and the lowest in blank control group(all P<0.05).Western blot results showed that relative expression levels of NLRP3,ASC,pro-Caspase-1,IL^-1βand IL^-18,thioredoxin and thioredoxin interacting protein in model group and blank vector group were significantly higher than those in silencing group,and were the lowest in blank control group on the 12th and 20th days(all P<0.05).Conclusion Up-regulation of NLRP3 inflammatory body and downstream inflammatory factors may be involved in occurrence of AU and may mediate ROS signaling pathway.

关 键 词：自身免疫性葡萄膜炎 活性氧簇 信号通路 核苷酸结合寡聚化结构域样受体 

分 类 号：R773[医药卫生—眼科]