Department of Ophthalmology, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
Invest Ophthalmol Vis Sci. 2014 May 15;55(6):3700-8. doi: 10.1167/iovs.14-14154.
To characterize the alterations of extracellular matrix proteins in Descemet's membranes (DM) of patients with late-onset Fuchs' corneal dystrophy (FCD) and to differentiate them from nonspecific alterations in pseudophakic bullous keratopathy (PBK).
Human DM-endothelial cell complexes were obtained from patients with late-onset FCD (n = 40), PBK (n = 6), and control eyes (n = 5). Gene expression profiles of endothelial cells were compared using a commercial real-time PCR array and quantitative real-time PCR assays for confirmation of differentially expressed genes. A total of 24 extracellular matrix proteins were also localized in cryosections of corneal specimens from FCD (n = 10), PBK (n = 4), and control eyes (n = 5) by immunohistochemistry.
Polymerase chain reaction array analysis revealed a significant upregulation of 27 out of 84 extracellular matrix-related genes including collagens, proteoglycans, glycoproteins, cell adhesion molecules, and matrix metalloproteinases in FCD specimens as compared to normal controls, which could be partly confirmed and quantified by real-time PCR. Comparative analysis of FCD and PBK specimens showed a significant and consistent FCD-specific upregulation of collagen types I, III, and XVI; fibronectin; agrin; clusterin; transforming growth factor beta-induced (TGFBI); and integrin α4 (3- to 18-fold, P < 0.05). Immunohistochemistry revealed an increased labeling of collagen (types III, VII, XV, XVI), agrin, fibulin-2, TGFBI, versican, and clusterin in the DM of FCD specimens compared to PBK specimens.
The findings provide evidence for a specific upregulation, production, and deposition of collagen types III and XVI, agrin, TGFBI, and clusterin in late-onset FCD and thus point to the importance of matrix alterations in the pathophysiology of FCD.
研究迟发性 Fuchs 角膜营养不良（FCD）患者的角膜后弹力层（Descemet's 膜，DM）中细胞外基质蛋白的变化，并将其与后发性白内障囊外摘出术后单纯大泡性角膜病变（PBK）的非特异性改变相区分。
从迟发性 FCD 患者（n=40）、PBK 患者（n=6）和对照眼（n=5）中获取 DM-内皮细胞复合物。采用商业实时 PCR 阵列和定量实时 PCR 检测对比分析内皮细胞的基因表达谱，以确认差异表达基因。应用免疫组织化学法在 FCD（n=10）、PBK（n=4）和对照眼（n=5）的角膜冷冻切片中定位 24 种细胞外基质蛋白。
PCR 阵列分析显示，与正常对照组相比，FCD 组织中有 27 种细胞外基质相关基因（包括胶原、蛋白聚糖、糖蛋白、细胞黏附分子和基质金属蛋白酶）显著上调，实时 PCR 部分证实并量化了这一结果。FCD 和 PBK 标本的对比分析显示，FCD 特异性的胶原 I、III、XVI、纤维连接蛋白、聚集蛋白、转化生长因子β诱导（TGFBI）和整合素α4 显著上调（3-18 倍，P<0.05）。免疫组织化学显示，与 PBK 标本相比，FCD 标本的 DM 中胶原（III、VII、XV、XVI）、聚集蛋白、纤维连接蛋白-2、TGFBI、玻璃体连接蛋白和聚集蛋白的标记增加。
这些发现为迟发性 FCD 中胶原 III 和 XVI、聚集蛋白、TGFBI 和聚集蛋白的特异性上调、产生和沉积提供了证据，因此提示基质改变在 FCD 病理生理学中的重要性。
