Barati Michelle T, Gould James C, Salyer Sarah A, Isaacs Susan, Wilkey Daniel W, Merchant Michael L
Kidney Disease Program, Department of Medicine, University of Louisville, Louisville, KY 40202, USA.
Kidney Disease Program, Department of Medicine, University of Louisville, Louisville, KY 40202, USA; Harvard Medical School, Boston, MA 02115, USA.
J Diabetes Res. 2016;2016:3537863. doi: 10.1155/2016/3537863. Epub 2015 Dec 29.
The effects of acute exposure to high glucose levels as experienced by glomerular mesangial cells in postprandial conditions and states such as in prediabetes were investigated using proteomic methods. Two-dimensional gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry methods were used to identify protein expression patterns in immortalized rat mesangial cells altered by 2 h high glucose (HG) growth conditions as compared to isoosmotic/normal glucose control (NG(⁎)) conditions. Unique protein expression changes at 2 h HG treatment were measured for 51 protein spots. These proteins could be broadly grouped into two categories: (1) proteins involved in cell survival/cell signaling and (2) proteins involved in stress response. Immunoblot experiments for a protein belonging to both categories, prohibitin (PHB), supported a trend for increased total expression as well as significant increases in an acidic PHB isoform. Additional studies confirmed the regulation of proteasomal subunit alpha-type 2 and the endoplasmic reticulum chaperone and oxidoreductase PDI (protein disulfide isomerase), suggesting altered ER protein folding capacity and proteasomal function in response to acute HG. We conclude that short term high glucose induces subtle changes in protein abundances suggesting posttranslational modifications and regulation of pathways involved in proteostasis.
采用蛋白质组学方法，研究了餐后状态下（如糖尿病前期）肾小球系膜细胞急性暴露于高葡萄糖水平的影响。二维凝胶电泳和基质辅助激光解吸电离飞行时间质谱法用于鉴定永生化大鼠系膜细胞在2小时高糖（HG）生长条件下与等渗/正常葡萄糖对照（NG(⁎)）条件相比所改变的蛋白质表达模式。在2小时HG处理下，对51个蛋白质斑点的独特蛋白质表达变化进行了测量。这些蛋白质大致可分为两类：（1）参与细胞存活/细胞信号传导的蛋白质，以及（2）参与应激反应的蛋白质。对属于这两类的一种蛋白质——抑制素（PHB）进行的免疫印迹实验支持了总表达增加的趋势以及酸性PHB同工型的显著增加。进一步的研究证实了蛋白酶体亚基α-2型以及内质网伴侣和氧化还原酶PDI（蛋白质二硫键异构酶）的调节，表明内质网蛋白质折叠能力和蛋白酶体功能在急性HG反应中发生了改变。我们得出结论，短期高糖会诱导蛋白质丰度的细微变化，提示蛋白质稳态相关途径的翻译后修饰和调节。
