A global proteome approach in uric acid stimulated human aortic endothelial cells revealed regulation of multiple major cellular pathways

•First study using proteomics to show UA dependent protein expression in HAEC•125 out of 771 proteins could be assigned to major cellular signaling pathways.•Ubiquitin-proteasome system is the most regulated pathway of UA.•In addition UA regulates eIF4 known to be essential for protein synthesis.

BackgroundUric acid (UA) has been identified as one major risk factor for cardiovascular diseases. Lowering of serum UA levels improves endothelial function. The present study investigates for the first time concentration-dependent effects of UA on human aortic endothelial cells (HAEC) and the cellular pathways involved in global proteomic analysis.MethodsThe concentration dependent effects of UA on HAEC were investigated by nanoLC-MS/MS and ingenuity pathway analysis to reveal putative cellular pathways. For verification of the identified pathways the abundance or activity of key proteins was measured using ELISA or Western blotting. NO production was quantified by confocal laser microscopy.ResultsWe identified ubiquitin-proteasome system (UPS) and eIF4 signaling as the major pathways regulated by UA. K-means clustering analysis revealed 11 additional pathways, of which NO, superoxide signaling and hypoxia were further analyzed. A complex regulatory network was detected demonstrating that 500 μmol/L UA, which is well above the concentration regarded as pathological in clinical settings, led to diminishing of NO bioavailability. In addition a UA-dependent downregulation of eIF4, an upregulation of UPS and an increase in HIF-1α were detected.ConclusionsHere we show for the first time, that increasing UA levels activate different sets of proteins representing specific cellular pathways important for endothelial function. This indicates that UA may alter far more pathways in HAEC than previously assumed. This regulation occurs in a complex manner depending on UA concentration. Further studies in knockout and overexpression models of the identified proteins are necessary to prove the correlation with endothelial dysfunction.