A Two-Stage Laser-Induced Mouse Model of Subretinal Fibrosis Secondary to Choroidal Neovascularization
A Two-Stage Laser-Induced Mouse Model of Subretinal Fibrosis Secondary to Choroidal Neovascularization
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DOI：
10.1167/tvst.9.4.3
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发表时间：
2020-03-01
影响因子：
3
通讯作者：
Xu, Heping
中科院分区：
文献类型：
--
作者：
Little, Karis;Llorian-Salvador, Maria;Xu, Heping
目的：方法：成年C57 BL/6 J小鼠每只眼接受3次激光烧伤以诱导脉络膜新生血管（CNV）。7天后，对每个新生血管病变进行第二次激光烧伤。传统的激光诱导CNV被用作对照。通过眼底成像、荧光素眼底血管造影、光学相干断层扫描（OCT）和免疫组织化学，在激光（p.l）治疗后10、20、30和40天监测病变。胶原-1（COL-1）、纤连蛋白、α-平滑肌肌动蛋白、F4/80、补体因子B（CF B）、补体成分3（C3）、转化生长因子-β（TGF-β）通过免疫荧光和逆转录聚合酶链反应检测视网膜和视网膜色素上皮/脉络膜中的TGF-β、成纤维细胞生长因子2（FGF 2）。在所有时间点，通过OCT和免疫组化，两阶段激光方案诱导的病变均显著大于传统激光CNV。共聚焦显微镜检测了两阶段激光方案术后30至40天病变中的COL-1（+）纤维和IBA 1（+）/CD 31（+）血管。在这个时间点，来自传统激光CNV的病变仅包含COL-1（+）纤维而不包含血管。与传统激光CNV损伤相比，在两阶段激光诱导损伤的视网膜中检测到更高水平的促炎基因（诱导型一氧化氮合酶（iNOS）、C3、CFB）和促纤维化基因（TGF-β、FGF 2和血管内皮生长因子）。在两阶段激光诱导的纤维化lesion.Conclusions：两阶段激光治疗诱导视网膜下纤维血管膜，持续超过40 days.Translational Relationship：该模型是一个有用的工具，研究黄斑纤维化的机制，在nAMD和测试抗纤维化药物。
Purpose: To develop a model that can recapitulate key features of macular fibrosis in neovascular age-related macular degeneration (nAMD).Methods: Adult C57BL/6J mice received three laser burns/eye to induce choroidal neovascularization (CNV). Seven days later, a second laser burn was directed to each of the neovascular lesions. Traditional laser-induced CNV was used as a control. Lesions were monitored at 10, 20, 30, and 40 days post-laser (p.l) treatment by fundus imaging, fundus fluorescein angiography, optical coherence tomography (OCT), and immunohistochemistry. The expression of collagen-1 (COL-1), fibronectin, alpha-smooth muscle actin, F4/80, complement factor B (CFB), Complement component 3 (C3), transforming growth factor-beta (TGF-beta), and fibroblast growth factor 2 (FGF2) in retina and retinal pigment epithelium/choroid was examined by immunofluorescence and reverse transcription polymerase chain reaction.Results: The two-stage laser protocol induced significantly larger lesions than the traditional laser-CNV by OCT and immunohistochemistry at all time points. Confocal microscopy detected COL-1(+) fibers and IBA1(+)/CD31(+) blood vessels in lesions from the two-stage laser protocol 30 to 40 days p.l. Lesions from traditional laser-CNV contain only COL-1(+) fibers but not blood vessels at this time point. Higher levels of proinflammatory (inducible nitric oxide synthase (iNOS), C3, CFB) and profibrotic (TGF-beta, FGF2, and vascular endothelial growth factor) genes were detected in the retinas from the two-stage laser-induced lesions compared with the traditional laser-CNV lesion. Higher number of infiltrating F4/80 macrophages was also observed in and around the two-stage laser-induced fibrotic lesion.Conclusions: The two-stage laser treatment induced subretinal fibrovascular membranes that persist over 40 days.Translational Relevance: The model is a useful tool to study the mechanism of macular fibrosis in nAMD and test antifibrotic drugs.