CCAAT/Enhancer-Binding Protein β Mediates Oxygen-Induced Retinal Neovascularization via Retinal Vascular Damage and Vascular Endothelial Growth Factor
CCAAT/Enhancer-Binding Protein β Mediates Oxygen-Induced Retinal Neovascularization via Retinal Vascular Damage and Vascular Endothelial Growth Factor
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CCAAT/增强子结合蛋白β通过视网膜血管损伤和血管内皮生长因子介导氧诱导的视网膜新生血管形成
DOI：
10.1155/2020/2789209
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发表时间：
2020-03-09
影响因子：
4.3
通讯作者：
Wu, Qiang
中科院分区：
文献类型：
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作者：
Li, Tingting;Cai, Xuan;Wu, Qiang
Objective.评价CCAAT/增强子结合蛋白β（C/EBP β）在氧诱导视网膜病变（OIR）模型中视网膜新生血管（RNV）中的作用。方法. OIR大鼠暴露于交替缺氧和远视条件下14天。然后，OIR大鼠被随机分配到接受玻璃体内注射靶向C/EBP β的shRNA慢病毒颗粒（LV.shC/EBP β）或对照颗粒（LV.shScrambled）的组。通过玻璃体内注射C/EBP β shRNA对OIR大鼠进行有效的转导，并通过视网膜荧光素-葡聚糖灌注、视网膜ADP染色和过碘酸-希夫（PAS）染色检测视网膜血管损伤和RNV的积聚。通过全视野闪光视网膜电图反应记录视网膜功能。采用逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测C/EBP β和血管内皮生长因子（VEGF）的mRNA和蛋白水平。蛋白质印迹分析检测p-C/EBP β的表达。通过免疫组化确定C/EBP β在视网膜中的表达位置。结果OIR大鼠C/EBP β和VEGF在mRNA和蛋白水平的表达均显著增加（P
Objective. To evaluate the role of CCAAT/enhancer-binding protein beta (C/EBP beta) in retinal neovascularization (RNV) in an oxygen-induced retinopathy (OIR) model. Methods. Rats with OIR were exposed to alternating hypoxic and hyperopic conditions for 14 days. Then, the rats with OIR were assigned randomly to groups that received intravitreal injections of either shRNA lentiviral particles targeting C/EBP beta (LV.shC/EBP beta) or control particles (LV.shScrambled). The effectiveness of transduction using intravitreal injection of C/EBP beta shRNA was examined in rats with OIR. The retinal vascular damage and accumulation of RNV were determined by retinal fluorescein-dextran perfusion, retinal ADPase staining, and periodic acid-Schiff (PAS) staining. Retinal function was recorded by electroretinogram responses to full-field light flashes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot analyses were used to measure mRNA and protein levels of C/EBP beta and vascular endothelial growth factor (VEGF). The expression of p-C/EBP beta was also examined by western blot analyses. The location of C/EBP beta expression in the retina was determined by immunohistochemistry. Results. In OIR rats, the expression levels of C/EBP beta and VEGF were significantly increased at both the mRNA and protein levels (P