Liu Gaoqin, Zhang Wenpeng, Xiao Yanhui, Lu Peirong
Department of Ophthalmology, the First Affiliated Hospital of Soochow University , Suzhou , China and.
Curr Eye Res. 2015 Sep;40(9):891-901. doi: 10.3109/02713683.2014.968934. Epub 2014 Oct 13.
To address the role of interferon-induced protein of 10 kDa (IP-10) in the course of corneal neovascularization (CrNV) in a mouse model of experimental corneal neovascularization.
BALB/c mice that were 7- to 8-week-old male were included in the study. Corneal injury was induced by NaOH. Mice were randomly divided into 2 groups of IP-10 and vehicle. The alkali-treated eyes received 5 μl of 5 μg/ml IP-10 dissolved in 0.2% sodium hyaluronate for IP-10-treated group, or 5 μl of 0.2% sodium hyaluronate for vehicle-treated group twice a day for 7 days immediately after the alkali injury. 2 weeks after alkali injury, corneas were removed and used for whole mount CD31 staining. The percentages of neovascularization on corneal photographs were examined with digital image analysis. In other experiments, at indicated time intervals, the corneas were removed. Angiogenic factor expression in the early phase after injury was quantified by real-time PCR and western blot. The VEGF expression in macrophages infiltrating into burned corneas was examined by Flow cytometry (FCM) and immunofluorescence. Tube formation and cell proliferation of human retinal endothelial cells (HRECs) were detected after being stimulated with IP-10 in vitro.
The mRNA and protein expression of IP-10 and C-X-C motif chemokine receptor 3 (CXCR3) was augmented after the alkali injury (p < 0.05). Compared with vehicle-treated mice, IP-10-treated mice exhibited reduced CrNV 2 weeks after injury, as evidenced by diminished CD31-positive areas (p < 0.05). Concomitantly, the intracorneal mRNA and protein expression enhancement of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) was lower in IP-10-treated mice than in vehicle-treated mice after injury (p < 0.05). Moreover, IP-10 inhibited HREC tube formation and proliferation in vitro.
IP-10-treated mice exhibited reduced alkali-induced CrNV through decreasing intracorneal VEGF and bFGF expression, and inhibiting endothelial cell proliferation and tube formation.
探讨10 kDa干扰素诱导蛋白（IP-10）在实验性角膜新生血管化小鼠模型的角膜新生血管化（CrNV）过程中的作用。
研究纳入7至8周龄的雄性BALB/c小鼠。通过氢氧化钠诱导角膜损伤。小鼠被随机分为IP-10组和载体组。碱处理后的眼睛，IP-10治疗组每天两次接受5 μl溶解于0.2%透明质酸钠中的5 μg/ml IP-10，载体治疗组每天两次接受5 μl 0.2%透明质酸钠，在碱损伤后立即进行，持续7天。碱损伤2周后，取出角膜用于全层CD31染色。用数字图像分析检查角膜照片上新生血管化的百分比。在其他实验中，在指定的时间间隔取出角膜。通过实时PCR和蛋白质印迹法定量损伤后早期血管生成因子的表达。通过流式细胞术（FCM）和免疫荧光检查浸润到烧伤角膜中的巨噬细胞中的VEGF表达。体外用IP-10刺激后检测人视网膜内皮细胞（HREC）的管形成和细胞增殖。
碱损伤后IP-10和C-X-C基序趋化因子受体3（CXCR3）的mRNA和蛋白表达增加（p < 0.05）。与载体治疗的小鼠相比，IP-10治疗的小鼠在损伤后2周时CrNV减少，CD31阳性区域减少证明了这一点（p < 0.05）。同时，损伤后IP-10治疗的小鼠角膜内血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）的mRNA和蛋白表达增强低于载体治疗的小鼠（p < 0.05）。此外，IP-10在体外抑制HREC管形成和增殖。
IP-10治疗的小鼠通过降低角膜内VEGF和bFGF表达以及抑制内皮细胞增殖和管形成，表现出碱诱导的CrNV减少。
