a Eye and ENT Hospital , Shanghai Medical College, Fudan University , Shanghai , People's Republic of China.
b Shanghai Key Laboratory of Visual Impairment and Restoration , Fudan University , Shanghai , People's Republic of China.
Curr Eye Res. 2018 Sep;43(9):1190-1198. doi: 10.1080/02713683.2018.1484144. Epub 2018 Jul 9.
To study retinal neovascularization (RNV) inhibition by intravitreal injections (IVs) of ranibizumab, sTie2 fusion protein (sTie2-Fc), and a combined therapy in an oxygen-induced retinopathy (OIR) model.
An OIR mouse model was used to simulate RNV in retinopathy of prematurity (ROP); and the effect of blocking the angiopoietin (Ang) and its receptor (Tie2) and the vascular endothelial growth factor (VEGF) and its receptor (VEGFR) signaling pathways was compared using an IV of sTie2-Fc (Ang inhibitor) and/or ranibizumab (aVEGF antagonist). The effects were assessed using fluorescein isothiocyanate (FITC)-dextran cardiac perfusion, isolectin B4 (IB4) staining with whole retinal mounting, and hematoxylin and eosin (HE) staining to count the endothelial cells (ECs) that broke through the internal limiting membrane (ILM). The mRNA and protein levels of VEGF-A, VEGFR-2, Ang1, Ang2, and Tie2 were also determined by reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analysis.
Compared with the control group injected with phosphate-buffered saline (PBS), all three experimental groups, ranibizumab, sTie2-Fc, and ranibizumab + sTie2-Fc, had a significant decrease in micro-vessel densities and neovascular clusters, and fewer ECs broke through the ILM (all p < 0.05). The non-perfusion areas decreased in both mono-treated groups, although the combined therapy had larger non-perfusion areas. All three treatments decreased the mRNA and protein levels of VEGFA, Ang1, and Tie2.
In this study, it was confirmed that blocking the Ang/Tie2 and/or VEGF/VEGFR pathways could inhibit RNV and decrease abnormal micro-vessel density; and the mono-blockage of Ang/Tie2 might cause a smaller non-perfusion area.
研究玻璃体内注射雷珠单抗、sTie2 融合蛋白（sTie2-Fc）和联合治疗对氧诱导视网膜病变（OIR）模型中视网膜新生血管（RNV）的抑制作用。
采用 OIR 小鼠模型模拟早产儿视网膜病变（ROP）中的 RNV，并通过玻璃体内注射 sTie2-Fc（Ang 抑制剂）和/或雷珠单抗（aVEGF 拮抗剂）比较阻断血管生成素（Ang）及其受体（Tie2）和血管内皮生长因子（VEGF）及其受体（VEGFR）信号通路的效果。通过荧光素异硫氰酸酯（FITC）-葡聚糖心脏灌注、全视网膜 mounting 的异硫氰酸荧光素-B4（IB4）染色和苏木精和伊红（HE）染色来评估内皮细胞（EC）突破内界膜（ILM）的数量。还通过逆转录聚合酶链反应（RT-PCR）和 Western blot 分析测定 VEGF-A、VEGFR-2、Ang1、Ang2 和 Tie2 的 mRNA 和蛋白水平。
与注射磷酸盐缓冲液（PBS）的对照组相比，雷珠单抗组、sTie2-Fc 组和雷珠单抗+sTie2-Fc 组的微血管密度和新生血管簇均显著降低，突破 ILM 的 EC 数量减少（均 p<0.05）。虽然单治疗组的非灌注区减少，但联合治疗组的非灌注区更大。三种治疗方法均降低了 VEGFA、Ang1 和 Tie2 的 mRNA 和蛋白水平。
在这项研究中，证实阻断 Ang/Tie2 和/或 VEGF/VEGFR 通路可抑制 RNV 并降低异常微血管密度；单阻断 Ang/Tie2 可能导致较小的非灌注区。
