神经元驱动的血管生成:NGF 在氧诱导的视网膜病变模型中的视网膜新生血管形成中的作用。

Neuronal-driven angiogenesis: role of NGF in retinal neovascularization in an oxygen-induced retinopathy model.

机构信息

State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.

出版信息

Invest Ophthalmol Vis Sci. 2010 Jul;51(7):3749-57. doi: 10.1167/iovs.09-4226. Epub 2010 Mar 5.

Abstract

PURPOSE. To evaluate the role of nerve growth factor (NGF) in retinal neovascularization in an oxygen-induced retinopathy (OIR) model. METHODS. The OIR model was established in C57BL/6J mice. NGF mRNA expression in retina was measured by quantitative real-time PCR. NGF expression in protein levels was evaluated by ELISA and immunostaining with NGF antibody. The effects of NGF on retinal neovascularization were evaluated by intravitreal injections of exogenous NGF and TrkA receptor inhibitor K252a, respectively, in an OIR model. Retinal neovascularization was measured by counting neovascular cell nuclei above the internal limiting membrane and by image quantification analysis in flat-mounted retinas perfused with fluorescein dextran. RESULTS. NGF mRNA in retina had significantly high expression at postnatal day (P)17 in the OIR model compared with normally developing mice. Similarly, ELISA and immunostaining assay showed significantly increased NGF expression in retina at P17 in OIR mice but no significant differences at P12 or P24 compared with normal controls. Exogenous NGF intraocular injection enhanced angiogenesis in the retina in the OIR model; however, injection with K252a, a high-affinity trkA receptor inhibitor, significantly decreased retinal neovascularization compared with that seen in the controls. CONCLUSIONS. NGF contributed to retinal neovascularization in the OIR model. Intravitreal injection with K252a, the trkA receptor inhibitor, reduced neovascularization, showing the potential therapeutic efficacy of NGF receptor inhibitor in OIR mice.

摘要

目的。评估神经生长因子(NGF)在氧诱导视网膜病变(OIR)模型中视网膜新生血管形成中的作用。

方法。在 C57BL/6J 小鼠中建立 OIR 模型。通过实时定量 PCR 测量视网膜中 NGF mRNA 的表达。通过 ELISA 和用 NGF 抗体进行免疫染色评估 NGF 在蛋白水平上的表达。通过分别在 OIR 模型中眼内注射外源性 NGF 和 TrkA 受体抑制剂 K252a 来评估 NGF 对视网膜新生血管形成的影响。通过计数内界膜上方的新生血管细胞核和对荧光素葡聚糖灌注的扁平视网膜进行图像定量分析来测量视网膜新生血管形成。

结果。与正常发育的小鼠相比,OIR 模型中出生后第 17 天(P)视网膜中 NGF mRNA 的表达明显升高。同样,ELISA 和免疫染色测定显示,OIR 小鼠的 P17 视网膜中 NGF 表达明显增加,但与正常对照相比,在 P12 或 P24 时无明显差异。外源性 NGF 眼内注射增强了 OIR 模型中视网膜的血管生成;然而,与对照组相比,高亲和力 trkA 受体抑制剂 K252a 的注射显著减少了视网膜新生血管形成。

结论。NGF 促进了 OIR 模型中的视网膜新生血管形成。眼内注射 trkA 受体抑制剂 K252a 可减少新生血管形成,表明 NGF 受体抑制剂在 OIR 小鼠中具有潜在的治疗效果。

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