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Proteomics reveals potential non-neuronal cholinergic receptor-effectors in endothelial cells.

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.

journal_name

Acta Pharmacol Sin

journal_title

Acta pharmacologica Sinica

authors

Zhang YY,Shen W,Zhang LC,Pan ZY,Long CL,Cui WY,Zhang YF,Wang H

doi

10.1038/aps.2014.38

subject

Has Abstract

pub_date

2014-09-01 00:00:00

pages

1137-49

issue

9

eissn

1671-4083

issn

1745-7254

pii

aps201438

journal_volume

35

pub_type

杂志文章

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