CCR7/p-ERK1/2/VEGF信号通路在氧诱导性视网膜病变小鼠模型中促进视网膜新生血管形成。
CCR7/p-ERK1/2/VEGF signaling promotes retinal neovascularization in a mouse model of oxygen-induced retinopathy.
作者信息
Yuan Lin-Hui, Chen Xiao-Long, Di Yu, Liu Mei-Lin
机构信息
Department of Ophthalmology, Shengjing Affiliated Hospital of China Medical University, Shenyang 110000, Liaoning Province, China.
出版信息
Int J Ophthalmol. 2017 Jun 18;10(6):862-869. doi: 10.18240/ijo.2017.06.06. eCollection 2017.
AIM
To investigate the role of CCR7/p-ERK1/2/VEGF signaling in the mouse model of oxygen-induced retinopathy (OIR).
METHODS
Neonatal C57BL/6J mice were evenly randomized into four groups: normoxia, OIR, OIR control (treated with scramble siRNA), and OIR treated (treated with CCR7 siRNA). Normoxia group was not specially handled. Postnatal day 7 (P7) mice in the OIR group were exposed to 75%±5% oxygen for 5d (P7-P12) and then maintained under normoxic conditions for 5d (P12-P17). Mice in the OIR control and OIR treated groups were given injections of scramble or CCR7 siRNA plasmid on P12 before returning to normoxic conditions for 5d (P12-P17). Retina samples were collected from all mice on P17, stained with adenosine diphosphatase (ADPase), and retinal neovascularization (RNV) was assessed. Retinas were also stained with hematoxylin and eosin (H&E) for RNV quantitation. The distribution and expression of CCR7, p-ERK1/2 and vascular endothelial growth factor (VEGF) were assessed immunohistochemistry, Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR).
RESULTS
High oxygen promoted retinal neovascularization (<0.05) and increased the number of endothelial nuclei in new vessels extending from the retina to the vitreous body; CCR7 promoted this process (<0.05). CCR7 and VEGF mRNA were expressed at higher levels in the OIR and OIR control groups than in the normoxia and OIR treated groups. CCR7, p-ERK1/2, and VEGF protein were expressed in the retinas of mice in the OIR and OIR control groups. Intravitreal injection of CCR7 siRNA significantly reduced CCR7, p-ERK1/2, and VEGF expression in the OIR mouse model (all <0.05). CCR7 significantly enhanced the neovascularization and non-perfusion areas in the OIR group (<0.05). CCR7 siRNA significantly reduced levels of p-ERK1/2 and VEGF as compared to OIR controls (<0.05).
CONCLUSION
These results suggest that CCR7/p-ERK 1/2/VEGF signaling plays an important role in OIR. CCR7 may be a potential target for the prevention and treatment of retinopathy of prematurity.
目的
研究CCR7/p-ERK1/2/VEGF信号通路在氧诱导视网膜病变（OIR）小鼠模型中的作用。
方法
将新生C57BL/6J小鼠均匀随机分为四组：常氧组、OIR组、OIR对照组（用乱序siRNA处理）和OIR处理组（用CCR7 siRNA处理）。常氧组不做特殊处理。OIR组出生后第7天（P7）的小鼠暴露于75%±5%的氧气中5天（P7-P12），然后在常氧条件下维持5天（P12-P17）。OIR对照组和OIR处理组的小鼠在P12时注射乱序或CCR7 siRNA质粒，然后回到常氧条件下5天（P12-P17）。在P17时收集所有小鼠的视网膜样本，用腺苷二磷酸酶（ADPase）染色，并评估视网膜新生血管形成（RNV）。视网膜也用苏木精和伊红（H&E）染色用于RNV定量。通过免疫组织化学、蛋白质印迹和定量实时聚合酶链反应（qRT-PCR）评估CCR7、p-ERK1/2和血管内皮生长因子（VEGF）的分布和表达。
结果
高氧促进视网膜新生血管形成（<0.05），并增加从视网膜延伸至玻璃体的新血管中内皮细胞核的数量；CCR7促进了这一过程（<0.05）。CCR7和VEGF mRNA在OIR组和OIR对照组中的表达水平高于常氧组和OIR处理组。CCR7、p-ERK1/2和VEGF蛋白在OIR组和OIR对照组小鼠的视网膜中表达。玻璃体内注射CCR7 siRNA可显著降低OIR小鼠模型中CCR7、p-ERK1/2和VEGF的表达（均<0.05）。CCR7显著增强了OIR组的新生血管形成和无灌注区域（<0.05）。与OIR对照组相比，CCR7 siRNA显著降低了p-ERK1/2和VEGF的水平（<0.05）。
结论
这些结果表明CCR7/p-ERK １/2/VEGF信号通路在OIR中起重要作用。CCR7可能是预防和治疗早产儿视网膜病变的潜在靶点。
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