Extracellular Matrix Alterations in Late-Onset Fuchs' Corneal Dystrophy
Extracellular Matrix Alterations in Late-Onset Fuchs' Corneal Dystrophy
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DOI：
10.1167/iovs.14-14154
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发表时间：
2014-06-01
影响因子：
4.4
通讯作者：
Kruse, Friedrich E.
中科院分区：
文献类型：
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作者：
Weller, Julia M.;Zenkel, Matthias;Kruse, Friedrich E.
目的。目的探讨迟发性Fuchs角膜营养不良（FCD）患者Descemet膜（DM）细胞外基质蛋白的改变，并将其与假性角膜大泡性病变（PBK）的非特异性改变进行区分。从迟发性FCD患者（n = 40）、PBK患者（n = 6）和对照眼（n = 5）中获得人dm -内皮细胞复合物。内皮细胞的基因表达谱比较使用商用实时PCR阵列和定量实时PCR检测，以确认差异表达基因。通过免疫组织化学方法，FCD （n = 10）、PBK （n = 4）和对照眼（n = 5）的角膜标本冷冻切片中也发现了24种细胞外基质蛋白。聚合酶链反应阵列分析显示，与正常对照相比，FCD标本中84个细胞外基质相关基因中有27个显著上调，包括胶原蛋白、蛋白聚糖、糖蛋白、细胞粘附分子和基质金属蛋白酶，这可以通过实时PCR部分证实和定量。FCD和PBK标本的对比分析显示，FCD特异性的I、III和XVI型胶原蛋白显著且一致上调；纤连蛋白;微笑的;clusterin;转化生长因子诱导（TGFBI）；整合素α 4（3 ~ 18倍，P < 0.05）。免疫组织化学显示，与PBK标本相比，FCD标本DM中胶原（III型，VII型，XV型，XVI型），agrin, fibulin-2， TGFBI， versican和clusterin的标记增加。该研究结果为迟发性FCD中III型和16型胶原蛋白、agrin、TGFBI和clusterin的特异性上调、产生和沉积提供了证据，从而指出了基质改变在FCD病理生理中的重要性。
PURPOSE. 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).METHODS. 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.RESULTS. 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 alpha 4 (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.CONCLUSIONS. 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.