氧诱导视网膜病变小鼠模型中参与p21调控的miRNA表达谱分析  

Study on the mechanism of miRNA expression profile and enrichment analysis on p21 in oxygen-induced retinopathy mice

作　　者：刘勃实 韩金栋 黄欣远 李辉 曹靖靖 梁景黎 李筱荣 东莉洁 Liu Boshi;Han Jindong;Huang Xinyuan;Li Hui;Cao Jingjing;Liang Jingli;Li Xiaorong;Dong Lijie(Tianjin Medical University Eye Hospital,Tianjin Medical University Eye Institute,Tianjin Medical University School of Optometry and Ophthalmology,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,国家眼耳鼻喉疾病临床医学研究中心天津市分中心,天津市视网膜功能与疾病重点实验室,天津300384

出　　处：《中华眼底病杂志》2022年第9期762-767,共6页Chinese Journal of Ocular Fundus Diseases

基　　金：天津市教委科研计划项目(2020KJ179)。

摘　　要：目的观察氧诱导视网膜病变(OIR)小鼠视网膜组织中miRNA的表达,筛选与p21及视网膜新生血管(RNV)形成相关的miRNA。方法实验研究。40只健康7日龄C57BL/6J小鼠随机分为正常组和OIR组,每组20只。OIR组构建氧诱导RNV模型,正常组不做任何处理。小鼠17日龄时,处死小鼠,视网膜荧光铺片观察小鼠RNV发生情况;光学显微镜下计数突破视网膜内界膜的血管内皮细胞核。取视网膜组织行miRNA芯片分析,检测正常组与OIR组之间差异表达的miRNA。所得差异miRNA靶基因分别进行基于基因注释(GO)和京都基因与基因组百科全书(KEGG)的富集分析;通过Targetscan、MiRanda、MicroT-CDS数据库比对筛选可能与p21有关的miRNA及通路。组间两两比较采用独立样本t检验。结果与正常组比较,OIR组小鼠视网膜无灌注区面积、RNV及突破视网膜内界膜的血管内皮细胞核数量增加,差异均有统计学意义(t=18.800、9.025,P<0.05)。与正常组比较,OIR组有统计学差异表达的miRNA 54个,其中,上调、下调者分别为47、7个。从3个数据库与所得差异miRNA的比对结果中共筛选到与p21相关miRNA 13个,按差异倍数从高到低分别为miR-7218-5p、miR-322-5p、miR-224-5p、miR-335-5p、miR-329-3p、miR-362-3p、miR-532-5p、miR-20b-5p、miR-20a-5p、miR-195a-5p、miR-423-5p、miR-497a-5p、miR-129-5p。差异miRNA靶基因富集分析,得到1112个GO条目及50条KEGG通路,其中50个GO条目和13条KEGG通路与p21相关。结论在OIR模型中共筛选出13个与P21相关的miRNA。Objective To observe the expression of miRNA in retinal tissue of mice with oxygen-induced retinopathy(OIR),and screen miRNAs related to p21 and retinal neovascularization(RNV)formation.Methods A experimental study.Forty healthy 7-day-old C57BL/6J mice were randomly divided into normal group and OIR group,with 20 mice in each group.The oxygen induced RNV model was constructed in the OIR group,and no treatment was performed in the normal group.At the age of 17 days,the mice were killed and the RNV of mice was observed by retinal fluorescence;the nuclei of vascular endothelium that broke through the inner limiting membrane of retina were counted under light microscope.The retinal tissues were taken for miRNA chip analysis to detect the differentially expressed miRNAs between the normal group and the OIR group.The resulting differential miRNA target genes were subjected to enrichment analysis based on gene annotation(GO)and Kyoto Encyclopedia of genes and genomes(KEGG);miRNAs and pathways that may be related to p21 were screened through Targetscan,MiRanda and MicroT-CDs database alignment.Independent sample t-test was used for pairwise comparison between groups.Results Compared with the normal group,the area of nonperfusion area,RNV and the number of vascular endothelial nuclei that broke through the inner limiting membrane of the retina in the OIR group increased significantly,differences were statistically significant(t=18.800,9.025;P<0.05).Compared with the normal group,there were 54 miRNAs that were statistically differentially expressed in the OIR group,of which 47 were up-regulated and 7 were down-regulated.A total of 13 miRNAs related to p21 were screened from the alignment results of the three databases with the obtained differential miRNAs.According to the difference multiples,they were miR-7218-5p,miR-322-5p,miR-224-5p,miR-335-5p,miR-329-3p,miR-362-3p,miR-532-5p,miR-20b-5p,miR-20a-5p,miR-195a-5p,miR-423-5p,miR-497a-5p,and miR-129-5p.Differential miRNA target gene enrichment analysis yielded 1112 go entries

关 键 词：视网膜新生血管化 微RNAS 周期素依赖激酶抑制剂p21 动物实验 

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