Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, 1 East Jianshe Road, Zhengzhou, 450052, China.
Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, 1 East Jianshe Road, Zhengzhou, 450052, China.
Exp Eye Res. 2019 Apr;181:232-239. doi: 10.1016/j.exer.2019.02.006. Epub 2019 Feb 6.
Proliferative vitreoretinopathy (PVR) is the leading cause of retinal detachment failure. The mechanism of PVR development is complex and still not completely elucidated. There are no proven methods for early prevention or clinical treatment. Retinal proteins are abnormally expressed during the entire PVR disease process. Due to the limitations of research methods and techniques, we do not fully understand the retinal protein changes in PVR. This proteomics study systemically analyzed and identified differential protein expression between retinas of PVR and non-PVR (normal) eyes. Retinal samples were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) coupled with mass spectrometry. Raw data were processed and analyzed by Maxquant software and then searched against the human UniProKB (201510) protein database. Differentially expressed proteins were selected and further validated in a human retinal pigment epithelial (RPE) cell line. The effects of dysregulated proteins on cell proliferation, apoptosis, and migration were studied. Systemic proteomics analysis identified several PVR-enriched proteins. The differentially expressed proteins were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation to find abnormal pathways involved in PVR. Retinal-specific ATP-binding cassette transporter (ABCA4) expression was one of the most increased proteins in PVR tissue. ABCA4 knockdown significantly reduced proliferation and affected the cell cycle in the human RPE cell line. ABCA4 knockdown also induced apoptosis and inhibited retinal cell migration. In conclusion, systemic proteomics analysis identified differentially expressed proteins in traumatic PVR, with ABCA4 being highly expressed. Disruption of ABCA4 expression induced apoptosis and inhibited cell proliferation and migration in a human RPE cell line.
增生性玻璃体视网膜病变（PVR）是视网膜脱离失败的主要原因。PVR 发展的机制很复杂，尚未完全阐明。目前尚无预防或临床治疗的有效方法。在整个 PVR 疾病过程中，视网膜蛋白异常表达。由于研究方法和技术的限制，我们不完全了解 PVR 中的视网膜蛋白变化。这项蛋白质组学研究系统地分析和鉴定了 PVR 与非 PVR（正常）眼的视网膜之间差异表达的蛋白质。将视网膜样品进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）与质谱法结合。通过 Maxquant 软件处理和分析原始数据，然后与人类 UniProKB（201510）蛋白质数据库进行搜索。选择差异表达蛋白并在人视网膜色素上皮（RPE）细胞系中进一步验证。研究了失调蛋白对细胞增殖、凋亡和迁移的影响。系统蛋白质组学分析鉴定了几种富含 PVR 的蛋白质。通过基因本体论（GO）和京都基因与基因组百科全书（KEGG）注释对差异表达蛋白进行分析，以发现 PVR 中涉及的异常途径。视网膜特异性 ATP 结合盒转运蛋白（ABCA4）的表达是 PVR 组织中增加最多的蛋白质之一。ABCA4 敲低显著降低了人 RPE 细胞系的增殖，并影响了细胞周期。ABCA4 敲低还诱导了细胞凋亡并抑制了视网膜细胞迁移。总之，系统蛋白质组学分析鉴定了外伤性 PVR 中的差异表达蛋白，其中 ABCA4 表达水平较高。ABCA4 表达的破坏诱导了人 RPE 细胞系中的细胞凋亡，并抑制了细胞增殖和迁移。
