Wang Xiao-Li, Fu Alex, Spiro Craig, Lee Hon-Chi
Departments of Internal Medicine, Mayo Clinic, 200 First Street SW, Rochester, Minnesota 55905, USA.
J Proteomics Bioinform. 2009 Jan 1;2:445. doi: 10.4172/jpb.1000104.
This study directly measured the relative protein levels in bovine aortic endothelial cells (BAEC) that were cultured for two weeks in normal (5 mM, NG) or high (22 mM, HG) glucose and then were subjected to laminar shear stress at 0 or 15 dynes/cm(2). Membrane preparations were labeled with one of the four isobaric tagging reagents (iTRAQ), followed by LC-MS/MS analysis. The results showed that HG and/or shear stress induced alterations in various membrane associated proteins involving many signaling pathways. While shear stress induced an increase in heat shock proteins and protein ubiquitination, which remained enhanced in HG, the effects of shear stress on the mechanosensing and protein phosphorylation pathways were altered by HG. These results were validated by Western blot analysis, suggesting that HG importantly modulates shear stress-mediated endothelial function.
本研究直接测量了在正常（5 mM，NG）或高糖（22 mM，HG）环境中培养两周，然后分别在0或15达因/平方厘米的层流切应力作用下的牛主动脉内皮细胞（BAEC）中的相对蛋白质水平。膜制剂用四种等压标记试剂（iTRAQ）之一进行标记，随后进行液相色谱-串联质谱（LC-MS/MS）分析。结果表明，高糖和/或切应力诱导了涉及许多信号通路的各种膜相关蛋白的改变。虽然切应力诱导热休克蛋白增加和蛋白质泛素化，且在高糖环境中仍保持增强，但高糖改变了切应力对机械传感和蛋白质磷酸化途径的影响。这些结果通过蛋白质印迹分析得到验证，表明高糖对切应力介导的内皮功能有重要调节作用。
