Proteomics reveals potential non-neuronal cholinergic receptor-effectors in endothelial cells |
| |
Authors: | Yuan-yuan Zhang Wei Shen Lian-cheng Zhang Zhi-yuan Pan Chao-liang Long Wen-yu Cui Yan-fang Zhang Hai Wang |
| |
Affiliation: | 1.Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China;2.Cardiovascular Drug Research Center, Thadweik Academy of Medicine, Beijing 100039, China |
| |
Abstract: | Aim:The non-neuronal acetylcholine system (NNAS) in endothelial cells participates in modulating endothelial function, vascular tone, angiogenesis and inflammation, thus plays a critical role in cardiovascular diseases. In this study, we used a proteomic approach to study potential downstream receptor-effectors of NNAS that were involved in regulating cellular function in endothelial cells.Methods:Human umbilical vein endothelial cells were incubated in the presence of acetylcholine, oxotremorine, pilocarpine or nicotine at the concentration of 10 μmol/L for 12 h, and the expressed proteins in the cells were separated and identified with two-dimensional electrophoresis (2-DE) and LC-MS. The protein spots with the largest changes were identified by LC-MS. Biowork software was used for database search of the peptide mass fingerprints.Results:Over 1200 polypeptides were reproducibly detected in 2-DE with a pH range of 3–10. Acetylcholine, oxotremorine, pilocarpine and nicotine treatment caused 16, 9, 8 and 9 protein spots, respectively, expressed differentially. Four protein spots were identified as destrin, FK506 binding protein 1A (FKBP1A), macrophage migration inhibitory factor (MIF) and profilin-1. Western blotting analyses showed that treatment of the cells with cholinergic agonists significantly decreased the expression of destrin, FKBP1A and MIF, and increased the expression of profilin-1.Conclusion:A set of proteins differentially expressed in endothelial cells in response to cholinergic agonists may have important implications for the downstream biological effects of NNAS. |
| |
Keywords: | non-neuronal acetylcholine system endothelial cell cholinergic agonist destrin FK506 binding protein 1A macrophage migration inhibitory factor profilin-1 proteomics cardiovascular disease |
|
|