Lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor inhibits retinal neovascularization in mice of oxygen-induced retinopathy_Chinese Journal of Ocular Fundus Diseases

Authors：

Huang Liangyu , Ke Yifeng , Lin Tingting , Bu Shaochong , Ren Xinjun , Jiao Mingfei , Wang Yong , Hu Liying , Wang Qiong , Hong Yaru , Li Xiaorong ,  Dong Lijie

Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, The College of Optometry & Ophthalmology, Tianjin 300384, China;
Huang Liangyu is working on the Department of Ophthalmology, The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300381, China;
Huang Liangyu and Ke Yifeng are contributed equally to the article;

Corresponding author：

Dong Lijie, Email: aitaomubang@126.com

Keywords：

Retinal neovascularization/prevention&control; Polypyrimidine traet-binding protein/drug effects; Lentivirus infections; NF-E2-related factor 2; Heme oxygenase (decyclizing); Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2020.01.012

Video：

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Objective To investigate the inhibitory effect of lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR).Methods One hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group, simple OIR model group, OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group), with 16, 32, 32 and 32 mice, respectively. When the mice were 7 days old, the mice in the normal control group were fed in a routine environment, and the mice in the OIR model group, Vec group and PSF group were established OIR model. The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1×10¹¹ TU/ml) at the age of 12 days. No injection was performed in the normal control group and simple OIR group. RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Western blot analysis was applied to detect the protein expression of Nrf2, HO-1 and PSF. Results Of the normal control group, simple OIR model group, Vec group and PSF group, the number of pre-retinal neovascular cell nuclei were 0.00, 14.36±5.50, 15.67±4.96, 8.13±2.09, the non-perfusion area were 0.00%, (35.71±2.81)%, (36.57±4.53)%, (15.33±4.75)%, respectively. The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87, 165.70; P＜0.05). Compared with the normal control group, there were more pre-retinal neovascular cell nucleis and larger non-perfusion area in the simple OIR model group and Vec group (P＜0.05). Compared with the simple OIR model group and Vec group, there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P＜0.05). Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2, HO-1 (F=53.66, 83.54) and protein expression of Nrf2, HO-1 and PSF (F=58.38, 52.69, 24.79) among 4 groups were significant (P＜0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P＜0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P＜0.05). model group and Vec group (P＜0.05).Conclusion Intravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.

Citation： Huang Liangyu, Ke Yifeng, Lin Tingting, Bu Shaochong, Ren Xinjun, Jiao Mingfei, Wang Yong, Hu Liying, Wang Qiong, Hong Yaru, Li Xiaorong, Dong Lijie. Lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor inhibits retinal neovascularization in mice of oxygen-induced retinopathy. Chinese Journal of Ocular Fundus Diseases, 2020, 36(1): 53-59. doi: 10.3760/cma.j.issn.1005-1015.2020.01.012 Copy

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1. 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.
2. 张哲, 刘竹青, 刘巨平, 等. 二甲双胍联合抗血管内皮生长因子药物治疗糖尿病视网膜病变的可能协同作用[J]. 中华眼底病杂志, 2018, 34(5): 453-457. DOI: 10.3760/cma.j.issn.1005-1015.2018.05.008.Zhang Z, Liu ZQ, Liu JP, et al. The synergistic effect of metformin and anti-vascular endothelial growth factor in the treatment of diabetic retinopathy[J]. Chin J Ocul Fundus Dis, 2018, 34(5): 453-457. DOI: 10.3760/cma.j.issn.1005-1015.2018.05.008.
3. 牛瑞, 东莉洁, 马腾, 等. 结缔组织生长因子重组干扰载体慢病毒颗粒的构建及其对视网膜血管内皮细胞内源性结缔组织生长因子表达的抑制作用[J]. 中华眼底病杂志, 2018, 34(6): 580-585. DOI: 10.3760/cma.j.issn.1005-1015.2018.06.011.Niu R, Dong LJ, Ma T, et al. Construction of connective tissue growth factor recombinant interference vector lentiviral particle and its inhibitory effect on endogenous connective tissue growth factor expression in retinal vascular endothelial cells[J]. Chin J Ocul Fundus Dis, 2018, 34(6): 580-585. DOI: 10.3760/cma.j.issn.1005-1015.2018.06.011.
4. 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.
5. Shao Y, Chen J, Dong LJ, et al. A protective effect of PPARα in endothelial progenitor cells through regulating metabolism[J]. Diabetes, 2019, 68(11): 2131-2142. DOI: 10.2337/db18-1278.
6. Shao Y, Chen J, Freeman W, et al. Canonical Wnt signaling promotes neovascularization through determination of endothelial progenitor cell fate via metabolic profile regulation[J]. Stem cells, 2019, 37(10): 1331-1343. DOI: 10.1002/stem.3049.
7. 田芳, 李文博, 黄亮瑜, 等. 聚嘧啶束结合蛋白相关剪接因子对过氧化氢诱导下视网膜色素上皮细胞凋亡的影响[J]. 中华眼底病杂志, 2018, 34(2): 159-163. DOI: 10.3760/cma.j.issn.1005-1015.2018.02.012.Tian F, Li WB, Huang LY, et al. The effect of polypyrimidine tract binding protein-associated splicing factor on hydrogen peroxide induced apoptosis of retinal pigment epithelial[J]. Chin J Ocul Fundus Dis, 2018, 34(2): 159-163. DOI: 10.3760/cma.j.issn.1005-1015.2018.02.012.
8. 高美子, 黄亮瑜, 东莉洁, 等. Krüppel样因子6对于TGF-β1诱导的晶状体上皮细胞纤维化的调控作用研究[J]. 中国中医眼科杂志, 2018, 28(1): 4-11. DOI: 10.13444/j.cnki.zgzyykzz.2018.01.002.Gao MZ, Huang LY, Dong LJ, et al. Regulation of Krüppel-like factor 6 on TGF-β1-induced fibrosis of lens epithelial cells[J]. Chinese Journal of Chinese Ophthalmology, 2018, 28(1): 4-11. DOI: 10.13444/j.cnki.zgzyykzz.2018.01.002.
9. 刘爱华, 高美子, 黄亮瑜, 等. 叉头框转录因子F2小发夹RNA对人眼小梁网细胞外基质蛋白表达的抑制作用[J]. 中华实验眼科杂志, 2019, 37(6): 405-410. DOI: 10.3760/cma.j.issn.2095-0160.2019.06.002.Liu AH, Gao MZ, Huang LY, et al. The inhibitory effect of FoxF2 shRNA on the expression of extracellular matrix of human trabecular meshwork[J]. Chin J Exp Ophthalmol, 2019, 37(6): 405-410. DOI: 10.3760/cma.j.issn.2095-0160.2019.06.002.
10. Tian F, Dong LJ, Zhou Y, et al. Rapamycin-induced apoptosis in HGF-stimulated lens epithelial cells by AKT/mTOR, ERK and JAK2/STAT3 pathways[J]. Int J Mol Sci, 2014, 15(8): 13833-13848. DOI: 10.3390/ijms150813833.
11. 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.
12. Wei R, Dong L, Xiao Q, et al. Engagement of Toll-like receptor 2 enhances interleukin (IL)-17(+) autoreactive T cell responses via p38 mitogen-activated protein kinase signalling in dendritic cells[J]. Clin Exp Immunol, 2014, 178(2): 353-363. DOI: 10.1111/cei.12405.
13. 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.
14. 滕贺, 黄亮瑜, 田芳, 等. 衰老标记蛋白30高表达对紫外线诱导人晶状体上皮细胞凋亡的影响[J]. 中华眼科杂志, 2017, 53(11): 835-841. DOI: 10.3760/cma.j.issn.0412-4081.2017.11.007.Teng H, Huang LY, Tian F, et al. Effects of SMP-30 overexpression on apoptosis of human lens epithelial cells induced by ultraviolet B irradiation[J]. Chin J Ophthalmol, 2017, 53(11): 835-841. DOI: 10.3760/cma.j.issn.0412-4081.2017.11.007.
15. Li W, Dong L, Ma M, et al. Preliminary in vitro and in vivo assessment of a new targeted inhibitor for choroidal neovascularization in age-related macular degeneration[J]. Drug Des Devel Ther, 2016, 10: 3415-3423. DOI: 10.2147/DDDT.S115801.
16. 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.
17. 田芳, 赵今稚, 滕贺, 等. Krüppel样因子6经活化转录因子4通路对晶状体上皮细胞凋亡的调控作用[J]. 中华实验眼科杂志, 2018, 36(3): 181-186. DOI: 10.3760/cma.j.issn.2095-0160.2018.03.005.Tian F, Zhao JZ, Teng H, et al. Regulation of Krüppel-like factor 6 via activating transcription factor 4 pathway to apoptosis of human lens epithelial cells[J]. Chin J Exp Ophthalmol, 2018, 36(3): 181-186. DOI: 10.3760/cma.j.issn.2095-0160.2018.03.005.
18. 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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Chinese Journal of Ocular Fundus Diseases

Lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor inhibits retinal neovascularization in mice of oxygen-induced retinopathy

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