Expression and analysis of miRNA in retinal tissue of oxygen-induced retinopathy mice_Chinese Journal of Ocular Fundus Diseases

Authors：

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;

Corresponding author：

Li Xiaorong, Email: lixiaorong@tmu.edu.cn

Keywords：

Retinal neovascularization/etiology; MicroRNAs; Animal experimentation

DOI：

10.3760/cma.j.cn511434-20191105-00361

Video：

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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.

Citation： Liu Boshi, Dong Lijie, Huang Liangyu, Huang Xinyuan, Liu Xun, Wang Qiong, Hong Yaru, Han Jindong, Li Xiaorong. Expression and analysis of miRNA in retinal tissue of oxygen-induced retinopathy mice. Chinese Journal of Ocular Fundus Diseases, 2020, 36(7): 544-550. doi: 10.3760/cma.j.cn511434-20191105-00361 Copy

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1. Dong L, Nian H, Shao Y, et al. PTB-associated splicing factor inhibits IGF-1-induced VEGF upregulation in a mouse model of oxygen-induced retinopathy[J]. Cell Tissue Res, 2015, 360(2): 233-243. DOI: 10.1007/s00441-014-2104-5.
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3. Shao Y, Dong LJ, Takahashi Y, et al. miRNA-451a regulates RPE function through promoting mitochondrial function in proliferative diabetic retinopathy[J]. Am J Physiol Endocrinol Metab, 2019, 316(3): 443-452. DOI: 10.1152/ajpendo.00360.2018.
4. Li WB, Ma MW, Dong LJ, et al. MicroRNA-34a targets notch1 and inhibits cell proliferation in glioblastoma multiforme[J]. Cancer Biol Ther, 2011, 12(6): 477-483. DOI: 10.4161/cbt.12.6.16300.
5. Wang P, Luo Y, Duan H, et al. MicroRNA 329 suppresses angiogenesis by targeting CD146[J]. Mol Cell Biol, 2013, 33(18): 3689-3699. DOI: 10.1128/MCB.00343-13.
6. 刘勃实, 东莉洁, 李筱荣, 等. 慢病毒介导的微小RNA-191对小鼠视网膜新生血管的抑制作用[J]. 中华眼底病杂志, 2019, 35(5): 475-479. DOI: 10.3760/cma.j.issn.1005-1015.2019.05.010.Liu BS, Dong LJ, Li XR, et al. miR-191 inhibits oxygen-induced retinal neovascularization in mice[J]. Chin J Ocul Fundus Dis, 2019, 35(5): 475-479. DOI: 10.3760/cma.j.issn.1005-1015.2019.05.010.
7. Wang XH, Chen L. MicroRNA-370 suppresses the retinal capillary endothelial cell growth by targeting KDR gene[J]. Bratisl Lek Listy, 2017, 118(4): 202-207. DOI: 10.4149/BLL_2017_040.
8. 田芳, 东莉洁, 周玉, 等. 重组腺相关病毒-多聚嘧啶序列结合蛋白相关剪接因子对氧诱导视网膜新生血管形成的抑制作用[J]. 中华眼底病杂志, 2014, 30(5): 504-508. DOI: 10.3760/cma.j.issn.1005-1015.2014.05.019.Tian F, Dong LJ, Zhou Y, et al. Inhibition of oxygen induced retinal neovascularization by recombinant adeno-associated virus-polypyrimidine tract-binding protein-associated splicing factor intraocular injection in mice[J]. Chin J Ocul Fundus Dis, 2014, 30(5): 504-508. DOI: 10.3760/cma.j.issn.1005-1015.2014.05.019.
9. Xing XL, Huang LY, Lv YJ, et al. DL-3-n-butylphthalide protected retinal Müller cells dysfunction from oxidative stress[J]. Curr Eye Res, 2019, 44(10): 1112-1120. DOI: 10.1080/02713683.2019.1624777.
10. 田芳, 胡博杰, 李文博, 等. 高表达多聚嘧啶序列结合蛋白相关剪接因子对糖基化终产物诱导下视网膜Müller细胞凋亡的影响[J]. 中华眼底病杂志, 2019, 35(1): 70-75. DOI: 10.3760/cma.j.issn.1005-1015.2019.01.015.Tian F, Hu BJ, Li WB, et al. Effects of polypyramidine tract binding protein-associated splicing factor overexpression on apoptosis of human Müller cells under advanced glycation end products treatment[J]. Chin J Ocul Fundus Dis, 2019, 35(1): 70-75. DOI: 10.3760/cma.j.issn.1005-1015.2019.01.015.
11. 邢小丽, 黄亮瑜, 苏睿虹, 等. 丁基苯酞对过氧化氢诱导下视网膜Müller细胞凋亡的影响[J]. 中华眼底病杂志, 2018, 34(5): 481-486. DOI: 10.3760/cma.j.issn.1005-1015.2018.05.014.Xing XL, Huang LY, Su RH, et al. Effect of dl-3-n-butylphthalide on apoptosis of retinal Müller cells induced by hydrogen peroxide[J]. Chin J Ocul Fundus Dis, 2018, 34(5): 481-486. DOI: 10.3760/cma.j.issn.1005-1015.2018.05.014.
12. 邢小丽, 黄亮瑜, 张哲, 等. 丁基苯酞对H₂O₂诱导下视网膜色素上皮细胞凋亡的保护作用[J]. 中华眼底病杂志, 2019, 35(5): 485-492. DOI: 10.3760/cma.j.issn.1005-1015.2019.05.011.Xing XL, Huang LY, Zhang Z, et al. Effects of butylphthalide on hydrogen peroxide induced retinal pigment epithelial cells injury[J]. Chin J Ocul Fundus Dis, 2019, 35(5): 485-492. DOI: 10.3760/cma.j.issn.1005-1015.2019.05.011.
13. 漆晨, 东莉洁, 乐毅, 等. 多聚嘧啶序列结合蛋白相关剪接因子对体外培养的视网膜色素上皮细胞磷脂酰肌醇3激酶/丝氨酸-苏氨酸蛋白激酶信号通路的调控作用[J]. 中华眼底病杂志, 2015(4): 363-367. DOI: 10.3760/cma.j.issn.1005-1015.2015.04.013.Qi C, Dong LJ, Yue Y, et al. The regulation of PTB-associated splicing factor on phosphatidylinositol 3 kinase/Akt signaling pathway in retinal pigment epithelial cells[J]. Chin J Ocul Fundus Dis, 2015(4): 363-367. DOI: 10.3760/cma.j.issn.1005-1015.2015.04.013.
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Expression and analysis of miRNA in retinal tissue of oxygen-induced retinopathy mice

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