title: Proteomics reveals potential non-neuronal cholinergic receptor-effectors in endothelial cells.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Zhang
authors.affiliation: Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.affiliation: Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Lian-cheng
authors.initials: LC
authors.lastname: Zhang
authors.affiliation: Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Zhi-yuan
authors.initials: ZY
authors.lastname: Pan
authors.affiliation: Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Chao-liang
authors.initials: CL
authors.lastname: Long
authors.affiliation: Cardiovascular Drug Research Center, Thadweik Academy of Medicine, Beijing 100039, China.
authors.forename: Wen-yu
authors.initials: WY
authors.lastname: Cui
authors.affiliation: Cardiovascular Drug Research Center, Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Yan-fang
authors.initials: YF
authors.lastname: Zhang
authors.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.
authors.forename: Hai
authors.initials: H
authors.lastname: Wang
mesh_terms: D002478:Cells, Cultured; D018679:Cholinergic Agonists / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D011950:Receptors, Cholinergic / Q000378:metabolism*; D012984:Software
chemical_list: D018679:Cholinergic Agonists; D020543:Proteome; D011950:Receptors, Cholinergic
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