Division of Diabetes, Metabolism and Endocrinology, Department of Internal Medicine, Showa University School of Medicine, Tokyo, Japan.
Biochem Biophys Res Commun. 2010 Aug 20;399(2):221-6. doi: 10.1016/j.bbrc.2010.07.058. Epub 2010 Jul 21.
Diabetes can lead to serious microvascular complications including proliferative diabetic retinopathy (PDR), the leading cause of blindness in adults. Recent studies using gene array technology have attempted to apply a hypothesis-generating approach to elucidate the pathogenesis of PDR, but these studies rely on mRNA differences, which may or may not be related to significant biological processes. To better understand the basic mechanisms of PDR and to identify potential new biomarkers, we performed shotgun liquid chromatography (LC)/tandem mass spectrometry (MS/MS) analysis on pooled protein extracts from neovascular membranes obtained from PDR specimens and compared the results with those from non-vascular epiretinal membrane (ERM) specimens. We detected 226 distinct proteins in neovascular membranes and 154 in ERM. Among these proteins, 102 were specific to neovascular membranes and 30 were specific to ERM. We identified a candidate marker, periostin, as well as several known PDR markers such as pigment epithelium-derived factor (PEDF). We then performed RT-PCR using these markers. The expression of periostin was significantly up-regulated in proliferative membrane specimens. Periostin induces cell attachment and spreading and plays a role in cell adhesion. Proteomic analysis by LC/MS/MS, which permits accurate quantitative comparison, was useful in identifying new candidates such as periostin potentially involved in the pathogenesis of PDR.
糖尿病可导致严重的微血管并发症，包括增生性糖尿病视网膜病变（PDR），这是成年人致盲的主要原因。最近使用基因芯片技术的研究试图采用产生假说的方法阐明 PDR 的发病机制，但这些研究依赖于 mRNA 差异，这些差异可能与重要的生物学过程有关，也可能无关。为了更好地了解 PDR 的基本机制并确定潜在的新生物标志物，我们对来自 PDR 标本新生血管膜的混合蛋白提取物进行了液相色谱（LC）/串联质谱（MS/MS）分析，并将结果与非血管性视网膜前膜（ERM）标本进行了比较。我们在新生血管膜中检测到 226 种不同的蛋白质，在 ERM 中检测到 154 种。在这些蛋白质中，102 种是新生血管膜特有的，30 种是 ERM 特有的。我们鉴定了候选标志物骨粘连蛋白（periostin）以及几种已知的 PDR 标志物，如色素上皮衍生因子（PEDF）。然后，我们使用这些标志物进行了 RT-PCR。在增生性膜标本中，periostin 的表达显著上调。Periostin 诱导细胞附着和扩展，并在细胞黏附中发挥作用。LC/MS/MS 的蛋白质组学分析可进行准确的定量比较，有助于鉴定潜在参与 PDR 发病机制的新候选物，如 periostin。
