title: Nicotine and cotinine stimulate secretion of basic fibroblast growth factor and affect expression of matrix metalloproteinases in cultured human smooth muscle cells.
abstract: OBJECTIVE
  We have recently shown that nicotine and its metabolite cotinine are mitogenic for smooth muscle cells in vitro. In the present study, we examined the effect of nicotine and cotinine on the production of growth factors and the expression of matrix metallo-proteinases in smooth muscle cells.
  
  
  METHODS
  Smooth muscle cells were harvested from human arteries and grown in culture. Subconfluent cultures were incubated for 24 hours in M199 containing 0.1% fetal bovine serum with or without nicotine or cotinine at concentrations ranging from 10(-9) mol/L to 10(-6) mol/L. The supernatants and cell lysates were assayed by enzyme-linked immunosorbent assay for basic fibroblast growth factor (bFGF), tumor necrosis factor alpha (TNF-alpha), platelet-derived growth factor AB (PDGF-AB), and transforming growth factor beta (TGF-beta). Matrix metalloproteinase expression was determined in subconfluent cultures incubated in albumin with or without nicotine or cotinine at 10(-8) mol/L and 10(-7) mol/L for 6, 12, 18, 24 and 36 hours. Northern blot analyses were performed with human cDNA probes for collagenase-1, stromelysin-1, gelatinase A, gelatinase B, and triose phosphate isomerase. Blots were quantified by phosphor-imaging techniques.
  
  
  RESULTS
  Both nicotine and cotinine stimulated the production and secretion of bFGF in a dose-dependent manner. PDGF, TNF-alpha, and TGF-beta secretions were not significantly affected by nicotine or cotinine. Collagenase was up-regulated by nicotine at 18 and 24 hours (4.5-fold to 5.8-fold) and by cotinine at 18 hours (from 5.0-fold to 29-fold). Stromelysin-1 was up-regulated by nicotine and cotinine at 12 and 18 hours (1.5-fold to 7.0-fold). Gelatinase A generally peaked at 12 hours and was up-regulated by both agents (2.0-fold to 6.5-fold).
  
  
  CONCLUSIONS
  Nicotine and cotinine enhanced the production of bFGF, a major mitogen for smooth muscle cells, and up-regulated the expression of several matrix metalloproteinases that are critical in cell migration. These data demonstrate mechanisms by which smoking may contribute to the development of intimal hyperplasia, atherosclerosis, and aneurysms.
authors.affiliation: Department of Surgery, State University of New York at Buffalo, USA.
authors.forename: C S
authors.initials: CS
authors.lastname: Carty
authors.forename: P D
authors.initials: PD
authors.lastname: Soloway
authors.forename: S
authors.initials: S
authors.lastname: Kayastha
authors.forename: J
authors.initials: J
authors.lastname: Bauer
authors.forename: B
authors.initials: B
authors.lastname: Marsan
authors.forename: J J
authors.initials: JJ
authors.lastname: Ricotta
authors.forename: M
authors.initials: M
authors.lastname: Dryjski
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D003367:Cotinine / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis*; D006801:Humans; D066298:In Vitro Techniques; D008666:Metalloendopeptidases / Q000096:biosynthesis*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009538:Nicotine / Q000494:pharmacology*; D013268:Stimulation, Chemical; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D016207:Cytokines; D016222:Fibroblast Growth Factor 2; D009538:Nicotine; D008666:Metalloendopeptidases; D003367:Cotinine