Establishment of a New Animal Model of Focal Subretinal Fibrosis That Resembles Disciform Lesion in Advanced Age-Related Macular Degeneration
Establishment of a New Animal Model of Focal Subretinal Fibrosis That Resembles Disciform Lesion in Advanced Age-Related Macular Degeneration
复制标题
DOI：
10.1167/iovs.10-5189
复制
发表时间：
2011-08-01
影响因子：
4.4
通讯作者：
Ishibashi, Tatsuro
中科院分区：
文献类型：
--
作者：
Jo, Young-Joon;Sonoda, Koh-Hei;Ishibashi, Tatsuro
目的.视网膜下纤维化导致视力损害，特别是如果病变在黄斑中，如在晚期年龄相关性黄斑变性中经常观察到的。渗出的白细胞形成异常的血管，引发局部炎症，伴有局部神经胶质增生和基质产生。本研究旨在建立局灶性视网膜下纤维化的动物模型。将富含巨噬细胞的腹膜渗出液细胞（佩奇）注射到C57 BL/6或MCP-1敲除（KO）小鼠的视网膜下腔中。7天后，视网膜下纤维化组织的大小通过在脉络膜平坦载玻片上的胶质细胞酸性蛋白（GFAP）阳性视网膜胶质细胞的粘附面积来评估。肌纤维母细胞的变化和胶原合成检测α-平滑肌肌动蛋白（α-SMA）和Masson三色染色的组织切片，分别。在与活化的巨噬细胞共培养期间，还检测了视网膜色素上皮（RPE）细胞上的α-SMA表达。7天后通过眼底镜观察PEC注射小鼠的视网膜下纤维组织。该组织由一个单调的低细胞密度区域组成，表达α-SMA和胶原蛋白合成。类固醇和抗氧化剂治疗可以减少残留的胶质细胞。因为PEC注射的MCP-1 KO小鼠显示较少的残留胶质细胞，不仅外源性巨噬细胞，而且内源性巨噬细胞被激活。巨噬细胞直接诱导体外培养的RPE细胞发生肌纤维化。活化的巨噬细胞在被置于视网膜下腔中时形成视网膜下纤维化并诱导RPE细胞中的肌纤维化变化。（Invest Ophthalmol维斯科学。2011; 52：6089-6095）DOI：10.1167/iovs.10-5189
PURPOSE. Subretinal fibrosis causes damage to visual acuity, especially if the lesion is in the macula, as is frequently observed in advanced age-related macular degeneration. Exudate leukocytes form abnormal vessels that initiate regional inflammation accompanied with local glial proliferation and matrix production. The purpose of this study was to establish an animal model of focal subretinal fibrosis.METHODS. Macrophage-rich peritoneal exudate cells (PECs) were injected into the subretinal space of C57BL/6 or MCP-1 knockout (KO) mice. Seven days later, the size of the subretinal fibrotic tissue was evaluated by the adherent area of glial fibrillary acidic protein (GFAP)-positive retinal glial cells on choroidal flat mounts. Myofibroblastic changes and collagen synthesis were detected by alpha-smooth muscle actin (alpha-SMA) and Masson trichrome staining of the histologic section, respectively. alpha-SMA expression was also examined on retinal pigment epithelium (RPE) cells during co-culture with activated macrophages.RESULTS. Subretinal fibrous tissue was observed by funduscopy in PEC-injected mice after 7 days. The tissue consisted of a monotonous, low-cell-density area that expressed alpha-SMA with collagen synthesis. Both steroid and antioxidant treatment can reduce residual glia. Because PEC-injected MCP-1 KO mice showed less residual glia, not only exogenous macrophages, but also intrinsic macrophages were activated. The macrophages directly induced myofibrotic changes in RPE cells in vitro.CONCLUSIONS. Activated macrophages form subretinal fibrosis when they are placed in the subretinal space and induce myofibrotic changes in RPE cells. (Invest Ophthalmol Vis Sci. 2011; 52: 6089-6095) DOI:10.1167/iovs.10-5189