重组IV型胶原α2(IV)NC1结构域是视网膜毛细血管内皮细胞增殖的有效调节剂，并可抑制视网膜前新生血管形成。

Recombinant alpha2(IV)NC1 domain of type IV collagen is an effective regulator of retinal capillary endothelial cell proliferation and inhibits pre-retinal neovascularisation.

作者信息

Coleman Gary, Gardiner Tom A, Boutaud Ariel, Stitt Alan W

机构信息

Centre for Vision Science, School of Biomedical Science, Queen's University Belfast, Royal Victoria Hospital, Grosvenor Road, Belfast, BT12 6BA, Northern Ireland, UK.

出版信息

Graefes Arch Clin Exp Ophthalmol. 2007 Apr;245(4):581-7. doi: 10.1007/s00417-006-0396-1. Epub 2006 Aug 17.

DOI:10.1007/s00417-006-0396-1

PMID:16915401

Abstract

BACKGROUND

A recombinant form of the alpha2(IV)NC1 domain of type IV collagen has been shown to have potent anti-angiogenic activity although this peptide has not been studied in the context of proliferative retinopathies. In the current investigation we examined the potential for alpha2(IV)NC1 to regulate retinal microvascular endothelial cell function using a range of in vitro and in vivo assay systems.

MATERIALS AND METHODS

alpha2(IV)NC1 at concentrations between 0.1 and 1 mug/ml was added to retinal microvascular endothelial cells (RMECs) followed by assessment of cell attachment, proliferation and survival. This agent was also tested within a novel in vitro three-dimensional retinal angiogenesis assay and the number of angiogenic sprouts quantified. alpha2(IV)NC1 was also delivered intra-vitreally to mice with oxygen-induced proliferative retinopathy (OIR) and neovascularisation evaluated in comparison with vehicle-treated controls.

RESULTS

RMECs treated with alpha2(IV)NC1 (0.1, 0.5 and 1 microg/ml) showed delayed attachment at 3 h post-seeding, although this deficit had been restored at the 6-h time point. BrdU assay of DNA replication revealed that confluent RMECs treated with alpha2(IV)NC1 showed no measurable response in comparison with vehicle-treated controls. By contrast, proliferation of sub-confluent RMECs was significantly reduced by alpha2(IV)NC1 at 0.5 microg/ml (P<0.01). alpha2(IV)NC1 also induced apoptosis in RMECs and inhibited angiogenesis of pre-existing retinal vascular networks in vitro (P<0.001). Intra-vitreal injection of alpha2(IV)NC1 in the OIR model significantly inhibited pre-retinal neovascularisation compared with vehicle-treated controls (P<0.001).

CONCLUSION

alpha2(IV)NC1 inhibits angiogenesis in the retinal microvasculature. This recombinant protein has potential for the treatment of neovascularisation in proliferative retinopathies.

摘要

背景

IV型胶原蛋白的重组α2(IV)NC1结构域已显示出具有强大的抗血管生成活性，尽管该肽尚未在增殖性视网膜病变的背景下进行研究。在当前的研究中，我们使用一系列体外和体内检测系统，研究了α2(IV)NC1调节视网膜微血管内皮细胞功能的潜力。

材料和方法

将浓度在0.1至1微克/毫升之间的α2(IV)NC1添加到视网膜微血管内皮细胞（RMECs）中，随后评估细胞附着、增殖和存活情况。该试剂还在一种新型的体外三维视网膜血管生成检测中进行了测试，并对血管生成芽的数量进行了量化。α2(IV)NC1也通过玻璃体腔内注射给予氧诱导增殖性视网膜病变（OIR）小鼠，并与载体处理的对照组相比评估新生血管形成情况。

结果

用α2(IV)NC1（0.1、0.5和1微克/毫升）处理的RMECs在接种后3小时显示附着延迟，尽管这种缺陷在6小时时间点已恢复。DNA复制的BrdU检测显示，与载体处理的对照组相比，用α2(IV)NC1处理的汇合RMECs没有可测量的反应。相比之下，0.5微克/毫升的α2(IV)NC1显著降低了亚汇合RMECs的增殖（P<0.01）。α2(IV)NC1还诱导RMECs凋亡，并在体外抑制预先存在的视网膜血管网络的血管生成（P<0.001）。与载体处理的对照组相比，在OIR模型中玻璃体腔内注射α2(IV)NC1显著抑制视网膜前新生血管形成（P<0.001）。