miR-142-5p靶向调控叉头转录蛋白O亚族3介导辅助性T细胞17细胞炎性反应促进自身免疫性葡萄膜炎的发展  

miR-142-5p target regulation of forkhead transcription protein O subfamily 3 mediateing helper T cell 17 cell inflammation reaction promotes the development of autoimmune uveitis

作　　者：李媛[1] 秦廷玉[2] 郭龙[1] 李淑珍[1] 侯习武[2] Li Yuan;Qin Tingyu;Guo Long;Li Shuzhen;Hou Xiwu(Department of Ophthalmology,Shangqiu First People's Hospital,Shangqiu 450000,China;Department of Ophthalmology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]商丘市第一人民医院眼科,476000 [2]郑州大学第一附属医院眼科,450053

出　　处：《中华眼底病杂志》2021年第7期533-541,共9页Chinese Journal of Ocular Fundus Diseases

基　　金：河南省高等学校重点科研项目计划(19A320059)。

摘　　要：目的观察实验性自身免疫性葡萄膜炎(EAU)模型大鼠CD4+T细胞中miR-142-5p及叉头转录蛋白O亚族3(FOXO3)的表达,初步探讨两者靶向关系以及对EAU的影响。方法健康雄性Lewis大鼠10只随机分为模型组、对照组。模型组大鼠以过继免疫方式诱导EAU动物模型。免疫后20 d,取对照组、模型组大鼠眼球和引流淋巴结提取CD4+T细胞,并分为对照组、模型组、mimic-阴性对照(NC)组、miR-142-5p-mimic组、小干扰(si)-NC组、si-FOXO3组进行体外实验。miR-142-5p-mimic组、si-FOXO3组分别转染miR-142-5p-mimic、si-FOXO3。健康雄性Lewis大鼠25只随机分为模型组、转染mimic-NC组、转染miR-142-5p-mimic组、转染si-NC组、转染si-FOXO3组,分别注射上述体外实验组细胞。免疫后4、8、12、16、20 d行裂隙灯显微镜检查并进行EAU评分;免疫后20 d,苏木精-伊红染色行组织病理学分级。实时荧光定量聚合酶链反应检测各组大鼠CD4+T细胞和眼组织中miR-142-5p、FOXO3 mRNA相对表达量及细胞培养上清液中辅助性T细胞17(Th17)标志物白细胞介素(IL)-17、IL-22、维甲酸相关孤儿受体γ(RORγ)mRNA相对表达量。生物信息学网站及双荧光素酶预测miR-142-5p与FOXO3的靶向关系。组间比较采用单因素方差分析或t检验。结果模型组大鼠均出现不同程度EAU症状,随时间延长,其症状加重。与对照组比较,模型组大鼠CD4+T细胞中miR-142-5p mRNA相对表达量升高,FOXO3 mRNA相对表达量下降,差异均有统计学意义(t=7.374、10.423,P=0.002、0.001)。与mimic-NC组比较,miR-142-5p-mimic组大鼠CD4+T细胞中miR-142-5p mRNA相对表达量上升,差异有统计学意义(t=6.540,P=0.003)。与模型组、mimic-NC组、si-NC组比较,miR-142-5p-mimic组、si-FOXO3组大鼠CD4+T细胞培养上清液中IL-17、IL-22、RORγmRNA相对表达量均显著增加,差异有统计学意义(F=26.110、6.292、5.269、55.660、10.490、11.430,P<0.05)。与转染mimic-NC组比较,转染miR-142-5p-mimicObjective To observe the expression of miR-142-5p and forkhead transcription protein O subgroup 3(FOXO3)in CD4+T cells of experimental autoimmune uveitis(EAU)model rats,and preliminarily explore the targeting relationship between the two and the effect on EAU impact.Methods Ten Lewis rats were randomly divided into model group and control group.Rats in the model group wree induced an EAU animal model by adoptive immunization.Twenty days after immunization,CD4+T cells were extracted from the eyeballs and draining lymph nodes of rats in the control group and model group,and divided into control group,model group,mimic-negative control(NC)group,miR-142-5p-mimic group,and small interference(si)-NC group,si-FOXO3 group for in vitro experiments.The miR-142-5p-mimic group and si-FOXO3 group were transfected with miR-142-5p-mimic and si-FOXO3,respectively.Twenty-five Lewis rats were randomly divided into model group,mimic-NC transfected group,miR-142-5p-mimic transfected group,si-NC transfected group,and si-FOXO3 transfected group.The above-mentioned in vitro experimental groups were injected with cells respectively.Slit lamp microscopy and EAU score were performed on 4,8,12,16,20 days after immunization;on 20 days after immunization,hematoxylin-eosin staining was performed for histopathological grading.Real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression of miR-142-5p and FOXO3 mRNA in CD4+T cells and eye tissues of rats in each group,and helper T cell 17(Th17)marker interleukin(IL)-17,IL-22,retinoic acid-related orphan receptor gamma(ROR gamma)relative expression level in the supernatant.Bioinformatics website and dual luciferase was used to predict the targeting relationship between miR-142-5p and FOXO3.One-way analysis of variance or t test was used for comparison between groups.Results All rats in the model group showed symptoms of EAU to varying degrees,and the symptoms became worse with time.Compared with the control group,the relative expression of miR-142-5p mR

关 键 词：葡萄膜炎 Th 17细胞 叉头转录因子类 miR-142-5p 

分 类 号：R773.9[医药卫生—眼科]