Aluksanasuwan Siripat, Khamchun Supaporn, Thongboonkerd Visith
Medical Proteomics Unit, Office for Research and Development; and Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital; and Center for Research in Complex Systems Science, Mahidol University, Bangkok, Thailand.
Proteomics. 2017 Sep;17(17-18). doi: 10.1002/pmic.201700151. Epub 2017 Aug 24.
Recently, several studies employed various proteomic approaches to define diabetes-induced changes in renal proteins. However, functional significance of those datasets in diabetic nephropathy remained unclear. We thus performed integrative proteome network analysis of such datasets followed by various targeted functional studies in distal renal tubular cells treated with high glucose (HG) (25 mM) compared to normal glucose (NG) (5.5 mM) and NG + mannitol (M) (5.5 + 19.5 mM). The data showed that at 96 h when cell proliferation/death, tight junction protein and β-/F-actin expression and organization, and transepithelial resistance remained unchanged, only HG caused increased levels of HSP90, HSP70, and HSP60, and increased accumulation of intracellular protein aggregates. In addition, HG also induced overproduction of intracellular ROS, decreased catalase level, increased level of oxidatively modified proteins, increased intracellular ATP level, and defective transepithelial Ca transport. However, both HG and M increased the levels of ubiquitinated proteins. Taken together, this study demonstrated significant perturbations of distal renal tubular cells induced by HG based on targeted functional studies guided by integrative proteome network analysis. These data may, at least in part, lead to better understanding of the pathogenic mechanisms of diabetic nephropathy.
最近，几项研究采用了各种蛋白质组学方法来确定糖尿病引起的肾脏蛋白质变化。然而，这些数据集在糖尿病肾病中的功能意义仍不清楚。因此，我们对这些数据集进行了综合蛋白质组网络分析，随后在高糖（HG）（25 mM）处理的远端肾小管细胞中进行了各种靶向功能研究，并与正常葡萄糖（NG）（5.5 mM）和NG + 甘露醇（M）（5.5 + 19.5 mM）进行比较。数据显示，在96小时时，当细胞增殖/死亡、紧密连接蛋白以及β-/F-肌动蛋白的表达和组织以及跨上皮电阻保持不变时，只有HG导致HSP90、HSP70和HSP60水平升高，以及细胞内蛋白质聚集体的积累增加。此外，HG还诱导细胞内活性氧的过量产生，降低过氧化氢酶水平，增加氧化修饰蛋白的水平，增加细胞内ATP水平，并导致跨上皮钙转运缺陷。然而，HG和M都增加了泛素化蛋白的水平。综上所述，本研究在综合蛋白质组网络分析指导下的靶向功能研究基础上，证明了HG对远端肾小管细胞有显著的扰动作用。这些数据可能至少部分有助于更好地理解糖尿病肾病的发病机制。
