Mass Spectrometry-Based Label Free Quantitative Proteomics to study the effect of 3-PO in endothelial cells

The rapid development of label-free quantitative proteomic techniques has provided fast and low-cost measurement of protein expression levels in complex biological samples. In addition, the development of highly reproducible nano-HPLC separation, high resolution mass spectrometer, and delicate computational tools has greatly improved the reliability and accuracy of label-free, comparative LC-MS/MS (1). Commercially available data processing software is able to automatically detect, match, and quantify peptides from hundreds of different LC-MS experiments simultaneously, which provides a high-throughput technique in the drug discovery process. These label-free quantitative approaches have provided powerful tools for analyzing protein changes in large-scale proteomics studies. This approach has been applied for the first time in this study to evaluate the effects of 3PO [3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one] at endothelial level, whose dysfunction has been associated with several cardiovascular diseases including atherosclerosis, acute myocardial infarction and chronic thromboembolic diseases. 3PO has been shown to inhibit the glycolytic flux partially and transiently and to reduce pathological angiogenesis in a variety of disease models (2). In addition, it has been shown that an inflammatory activation of monocytes/macrophages, via Toll-like receptor ligands or pro-inflammatory cytokines, switches the endothelial cell (EC) metabolism from oxidative.