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,筛选与视网膜新生血管(RNV)有关的miRNA。方法80只健康7日龄C57BL/6J小鼠随机分为对照组和OIR组,每组40只。OIR组小鼠构建OIR模型,对照组小鼠不做任何处理。小鼠17日龄时,做视网膜荧光铺片,荧光显微镜下观察小鼠RNV发生情况;做视网膜病理切片HE染色,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,检测对照组与OIR组之间差异表达的miRNA,并行PCR验证。所得差异miRNA靶基因和表达谱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。组间两两比较采用独立样本t检验。结果荧光显微镜及光学显微镜观察发现,对照组小鼠视网膜未见无灌注区、新生血管及突破视网膜内界膜的血管内皮细胞核;OIR组小鼠视网膜可见大量无灌注区、新生血管及突破视网膜内界膜的血管内皮细胞核。两组小鼠突破视网膜内界膜的血管内皮细胞核数量比较,差异有统计学意义(t=9.025,P<0.05)。miRNA芯片分析结果显示,与对照组比较,OIR组有54个miRNA发生了具有统计学差异的表达。其中,上调者47个,下调者7个。miRNA差异表达倍数大于1.25倍者23个,小于0.75倍者5个。PCR验证结果显示,差异表达倍数最高的5个miRNA表达趋势与芯片分析结果一致,差异均有统计学意义(P<0.05)。GO分析共得到1112个差异有统计学意义的条目(P<0.05);KEGG分析共得到65个差异有统计学意义的条目(P<0.05)。结论OIR小鼠视网膜组织中miRNA较正常小鼠视网膜组织有54个表达差异显著的miRNA;表达差异的miRNA可能不同程度参与了RNV的发生发展。
Objective To analyze the expression of miRNA involved in regulating retinal neovascularizationin in retinal tissue of oxygen-induced retinopathy(OIR)mice.Methods Eighty healthy C57BL/6J mice were randomly divided into control group and OIR group at postnatal day 7(P7).Control group were not received any treatment and then exposed to room air.The OIR group was exposed to(75±2)%oxygen and then under room air at P12.Mice of all groups were euthanized at P17.Retinal neovasculation(RNV)was evaluated by counting the number of pre-retinal neovascular cells and analysing no perfusion area by immunofluorescent staining of the mouse retina.Total RNA was extracted from retinal tissue,and miRNA microarrays was performed to identify differentially expressed miRNA in the two groups.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis showed differential microRNA.Results Compared with the control group,the retinal neovascular tufts and the no perfusion area were both significantly smaller than those in OIR group.The number of pre-retinal neovascular cell nuclei in retinas from control group were obviously lower than those in the retinas from OIR group(t=9.025,P<0.05).MiRNA microarray analysis showed that 54 miRNA in OIR group showed statistically different expression in control group,47 miRNA were up-regulated and 7 miRNA were down-regulated.The results of PCR were consistent with the trend of microarray.In GO analysis,1112 items were significantly different(P<0.05),and 65 items were significantly different in KEGG analysis of expression profile(P<0.05).Conclusions The miRNA expression in retinal tissue of OIR mice is different from that of normal mice,and these miRNA may be involved in the development of RNV.There are 54 miRNA expression differences in retinal tissue of OIR compared with normal mouse retinal tissue.
作者
刘勃实
东莉洁
黄亮瑜
黄欣远
刘勋
王琼
洪雅茹
韩金栋
李筱荣
Liu Boshi;Dong Lijie;Huang Liangyu;Huang Xinyuan;Liu Xun;Wang Qiong;Hong Yaru;Han Jindong;Li Xiaorong(Tianjin Key Laboratory of Retinal Functions and Diseases,Tianjin International Joint Research and Development Centre of Ophthalmology and Vision Science,Eye Institute and School of Optometry,Tianjin Medical University Eye Hospital,Tianjin 300384,China)
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2020年第7期544-550,共7页
Chinese Journal of Ocular Fundus Diseases
基金
天津市教委科研计划项目(2017KJ215)。
