Elsevier

Surgery

Volume 127, Issue 1, January 2000, Pages 72-78
Surgery

Original Communications
Nicotine induces platelet-derived growth factor release and cytoskeletal alteration in aortic smooth muscle cells

https://doi.org/10.1067/msy.2000.102422Get rights and content

Abstract

Background: Cigarette smoking is implicated in atherosclerotic plaque formation, but the role of nicotine in this process is not completely understood. The release of platelet-derived growth factor (PDGF) by the bovine aortic smooth muscle cell (SMC) after nicotine administration at a concentration similar to that ingested by active and passive smokers and the role of PDGF in SMC cytoskeletal modification were studied. Methods: SMC, harvested with enzymatic digestion from calf aorta, were stimulated in a serum-free medium for 72 hours with (-)-nicotine (from 6 × 10-4 mol/L to 6 × 10-8 mol/L). The release of PDGF was assessed by inhibition antibody-binding assay and confirmed by Western blotting. Mitogenic activity of nicotine on SMCs was also determined. The SMC cytoskeleton was studied with specific antibodies anti-α-actin fibers, anti-vimentin, and anti-β-tubulin, and the modification induced by PDGF was assessed by blocking PDGF activity with specific antibodies. Results: The greatest PDGF release (1.24 ± 0.14 ng/104 cells vs control 0.43 ± 0.07 ng/104 cells) was noted at a (-)-nicotine concentration of 6 × 10-7 mol/L (P <.001). The addition of monoclonal antibody anti-PDGF decreased the tritiated thymidine uptake of SMCs exposed to (-)-nicotine compared with the control (29% vs 5% - P <.001). SMCs exposed to (-)-nicotine concentration of 6 × 10-7 mol/L and 6 × 10-8 mol/L had a significant alteration in the expression of α-actin fibers, vimentin, and β-tubulin compared with control. The administration of antibody anti-PDGF in the culture medium reversed cytoskeletal alteration. Conclusions: Nicotine enhanced the release of platelet-derived growth, which in turn caused an alteration in cytoskeletal organization. (Surgery 2000;127:72-8)

Section snippets

Cell culture

Smooth muscle cells were obtained from bovine thoracic aortas by collagenase digestion.22 The cells were seeded into 25 cm2 flasks (Falcon, Becton Dickinson Labware, Franklin Lakes, NJ) in a medium consisting of Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS) and antibiotics (penicillin 100 IU/mL, streptomycin 100 μg/mL, and gentamycin 200 μg/mL). The cultures were kept at 37°C in an atmosphere of 5% CO2 in air. The medium was changed after 1 day and

PDGF assay in the conditioned media

SMCs stimulated with (-)-nicotine released more PDGF than the control (P <.05; Table).

Table. Release of PDGF by SMCs (ng/104 cells)

(-)-Nicotine concentrationsPDGF
00.43 ± 0.07
6 × 10-8 mol/L1.00 ± 0.14
6 × 10-7 mol/L1.24 ± 0.14
6 × 10-6 mol/L1.00 ± 0.12
6 × 10-5 mol/L0.87 ± 0.10
6 × 10-4 mol/L0.61 ± 0.07
The release of PDGF was augmented in a dose-dependent manner with the increase of (-)-nicotine concentration from 6 × 10-8 mol/L to 6 × 10-7 mol/L. At higher (-)-nicotine concentrations (from 6 × 10-6

Discussion

Epidemiological studies have established a strong correlation between cigarette smoking and the development and progression of atherosclerosis, myocardial infarction, vascular graft failures, and death from coronary artery disease. Smokers have a 2.5-fold increase in the incidence of coronary heart disease, a 1.5-fold increase in stroke mortality, and an even greater increase in peripheral arterial occlusive disease as compared with nonsmokers.24 However, there is a paucity of information

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