Yohannes Elizabeth, Chang Jinsook, Christ George J, Davies Kelvin P, Chance Mark R
Case Center for Proteomics, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Mol Cell Proteomics. 2008 Jul;7(7):1270-85. doi: 10.1074/mcp.M700563-MCP200. Epub 2008 Mar 12.
Protein expression profiles in rat bladder smooth muscle were compared between animal models of streptozotocin-induced diabetes mellitus (STZ-DM) and age-matched controls at 1 week and 2 months after induction of hyperglycemia with STZ treatment. At each time point, protein samples from four STZ-DM and four age-matched control rat bladder tissues were prepared independently and analyzed together across multiple DIGE gels using a pooled internal standard sample to quantify expression changes with statistical confidence. A total of 100 spots were determined to be significantly changing among the four experimental groups. A subsequent mass spectrometry analysis of the 100 spots identified a total of 56 unique proteins. Of the proteins identified by two-dimensional DIGE/MS, 10 exhibited significant changes 1 week after STZ-induced hyperglycemia, whereas the rest showed differential expression after 2 months. A network analysis of these proteins using MetaCore suggested induction of transcriptional factors that are too low to be detected by two-dimensional DIGE and identified an enriched cluster of down-regulated proteins that are involved in cell adhesion, cell shape control, and motility, including vinculin, intermediate filaments, Ppp2r1a, and extracellular matrix proteins. The proteins that were up-regulated include proteins involved in muscle contraction (e.g. Mrlcb and Ly-GDI), in glycolysis (e.g. alpha-enolase and Taldo1), in mRNA processing (e.g. heterogeneous nuclear ribonucleoprotein A2/B1), in inflammatory response (e.g. S100A9, Annexin 1, and apoA-I), and in chromosome segregation and migration (e.g. Tuba1 and Vil2). Our results suggest that the development of diabetes-related complications in this model involves the down-regulation of structural and extracellular matrix proteins in smooth muscle that are essential for normal muscle contraction and relaxation but also induces proteins that are associated with cell proliferation and inflammation that may account for some of the functional deficits known to occur in diabetic complications of bladder.
在链脲佐菌素诱导的糖尿病（STZ-DM）动物模型与年龄匹配的对照之间，比较了用STZ处理诱导高血糖后1周和2个月时大鼠膀胱平滑肌中的蛋白质表达谱。在每个时间点，分别制备来自4个STZ-DM和4个年龄匹配对照大鼠膀胱组织的蛋白质样品，并使用混合内标样品在多个DIGE凝胶上一起分析，以通过统计学置信度定量表达变化。在四个实验组中，共确定100个斑点有显著变化。随后对这100个斑点进行质谱分析，共鉴定出56种独特蛋白质。通过二维DIGE/MS鉴定的蛋白质中，10种在STZ诱导高血糖后1周出现显著变化，其余的在2个月后显示差异表达。使用MetaCore对这些蛋白质进行网络分析，提示诱导了二维DIGE检测不到的转录因子，并鉴定出一个下调蛋白质的富集簇，这些蛋白质参与细胞粘附、细胞形状控制和运动，包括纽蛋白、中间丝、Ppp2r1a和细胞外基质蛋白。上调的蛋白质包括参与肌肉收缩（如Mrlcb和Ly-GDI）、糖酵解（如α-烯醇化酶和Taldo1）、mRNA加工（如不均一核核糖核蛋白A2/B1）、炎症反应（如S100A9、膜联蛋白1和载脂蛋白A-I）以及染色体分离和迁移（如微管蛋白α1和绒毛蛋白2）的蛋白质。我们的结果表明，该模型中糖尿病相关并发症的发展涉及平滑肌中结构和细胞外基质蛋白的下调，这些蛋白对正常肌肉收缩和舒张至关重要，但也诱导了与细胞增殖和炎症相关的蛋白质，这可能解释了膀胱糖尿病并发症中已知出现的一些功能缺陷。
