Hirahara Shuichiro, Nozaki Miho, Ohbayashi Masaharu, Hasegawa Norio, Ozone Daisuke, Ogura Yuichiro
Department of Ophthalmology and Visual Science, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Ophthalmic Res. 2017;58(1):56-66. doi: 10.1159/000463238. Epub 2017 Apr 5.
To investigate the association between retinal neovascularization and the CC chemokine receptor-3 (CCR3) in a mouse model of oxygen-induced retinopathy (OIR).
An OIR model in C57BL/6J mice was used as a retinal neovascularization model. An enzyme-linked immunosorbent assay was performed to evaluate the chronological change in vascular endothelial growth factor A (VEGF-A) and eotaxin expressions. CCR3 and VEGF subtype expression in the retina was examined using real-time RT-PCR, and CCR3, eotaxin, VEGF-A, and CD31 expression was examined immunohistochemically. A CCR3 neutralizing antibody (Ab) was injected into the vitreous humor on both postnatal days 12 (P12) and 14 (P14). Retinal neovascularizations were quantified by measurement of the percentages of neovascular area.
The mean eotaxin and VEGF-A protein level was significantly downregulated at P10 and P12 and was significantly upregulated at P14 and P17 (p < 0.05). CCR3 mRNA expression was significantly upregulated at P12 (p < 0.05). VEGF164 mRNA expression was significantly upregulated at P14 (p < 0.05). The areas of vaso-obliteration and neovascularization were significantly suppressed in anti-CCR3 Ab-treated eyes (p < 0.05). Anti-CCR3 Ab treatment suppressed VEGF and eotaxin but not monocyte chemoattractant protein-1. And VEGF 164 mRNA but not VEGF120 mRNA was suppressed by anti-CCR3 Ab treatment.
The present data suggest that anti-CCR3 treatment can suppress retinal neovascularization. Anti-CCR3 treatment may have potential as a new therapy for retinopathies with retinal neovascularization such as diabetic retinopathy and retinopathy of prematurity.
在氧诱导性视网膜病变（OIR）小鼠模型中研究视网膜新生血管与CC趋化因子受体3（CCR3）之间的关联。
将C57BL/6J小鼠的OIR模型用作视网膜新生血管模型。进行酶联免疫吸附测定以评估血管内皮生长因子A（VEGF-A）和嗜酸性粒细胞趋化因子表达的时间变化。使用实时逆转录聚合酶链反应检测视网膜中CCR3和VEGF亚型的表达，并通过免疫组织化学检测CCR3、嗜酸性粒细胞趋化因子、VEGF-A和CD31的表达。在出生后第12天（P12）和第14天（P14）将CCR3中和抗体（Ab）注入玻璃体腔。通过测量新生血管面积的百分比对视网膜新生血管进行定量。
嗜酸性粒细胞趋化因子和VEGF-A蛋白的平均水平在P10和P12时显著下调，在P14和P17时显著上调（p<0.05）。CCR3 mRNA表达在P12时显著上调（p<0.05）。VEGF164 mRNA表达在P14时显著上调（p<0.05）。抗CCR3 Ab治疗的眼睛中血管闭塞和新生血管的面积显著受到抑制（p<0.05）。抗CCR3 Ab治疗抑制了VEGF和嗜酸性粒细胞趋化因子，但未抑制单核细胞趋化蛋白-1。抗CCR3 Ab治疗抑制了VEGF 164 mRNA，但未抑制VEGF120 mRNA。
目前的数据表明抗CCR3治疗可抑制视网膜新生血管。抗CCR3治疗可能具有作为糖尿病性视网膜病变和早产儿视网膜病变等伴有视网膜新生血管的视网膜病变新疗法的潜力。
