title: Nicotine improves the functional activity of late endothelial progenitor cells via nicotinic acetylcholine receptors.
abstract: The aim of this study is to investigate whether nicotinic acetylcholine receptors (nAChRs) are involved in the modulation of functional activity of late endothelial progenitor cells (EPCs) induced by nicotine. Total mononuclear cells (MNCs) were isolated from human umbilical cord blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture plates. Late EPCs were positive for 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine-labeled acetylated low-density lipoprotein (DiI-acLDL) uptake and fluorescein-isothiocyanate-conjugated Ulex europaeus agglutinin lectin (UEA-1) binding. Expression of von Willbrand factor (vWF), kinase insert domain receptor (KDR), and α7 nAChR was detected by indirect immunofluorescence staining. Late EPCs of 3-5 passages were treated for 32 h with either vehicle or nicotine with or without pre-incubation of nAChR antagonism, mecamylamine, or α-bungarotoxin. The viability, migration, and in vitro vasculogenesis activity of late EPCs were assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, modified Boyden chamber assay, and in vitro angiogenesis assay, respectively. Late EPCs adhesion assay was performed by replating cells on fibronectin-coated plates, and then adherent cells were counted. Incubation with 10 nmol/L nicotine enhanced viable, migratory, adhesive, and in vitro vasculogenesis capacity of late EPCs. The effect of nicotine on late EPCs can be attenuated by mecamylamine or α-bungarotoxin. In conclusion, nicotine improves the functional activity of late EPCs via nAChRs.
authors.affiliation: Department of Cardiology, the First Affiliated Hospital, Shantou University Medical College, Guangdong, China.
authors.forename: Min
authors.initials: M
authors.lastname: Yu
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.forename: Kaihong
authors.initials: K
authors.lastname: Yi
authors.forename: Libiao
authors.initials: L
authors.lastname: Wu
authors.forename: Xuerui
authors.initials: X
authors.lastname: Tan
mesh_terms: D002038:Bungarotoxins / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D048430:Cell Shape; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D007231:Infant, Newborn; D008464:Mecamylamine / Q000494:pharmacology; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009538:Nicotine / Q000494:pharmacology*; D018722:Nicotinic Agonists / Q000494:pharmacology*; D018733:Nicotinic Antagonists / Q000494:pharmacology; D011978:Receptors, Nicotinic; D013234:Stem Cells / Q000187:drug effects* / Q000502:physiology
chemical_list: D002038:Bungarotoxins; D018722:Nicotinic Agonists; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; D008464:Mecamylamine; D009538:Nicotine