Nicotinic and PDGF-receptor function are essential for nicotine-stimulated mitogenesis in human vascular smooth muscle cells
✍ Scribed by Ivo A. Pestana; Roberto I. Vazquez-Padron; Abdelouahab Aitouche; Si M. Pham
- Publisher
- John Wiley and Sons
- Year
- 2005
- Tongue
- English
- Weight
- 211 KB
- Volume
- 96
- Category
- Article
- ISSN
- 0730-2312
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✦ Synopsis
Abstract
Cigarette smoking is implicated in the formation of occlusive vascular diseases. Nicotine's role in this process is incompletely understood. Nicotine's effect on human aortic vascular smooth muscle cells (HaVSMC) and the role of the nicotinic receptor (nAChR), platelet‐derived growth factor (PDGF), and the PDGF‐receptor (PDGF‐R) in this response were studied. Nicotine's mitogenic effect was characterized by three methods: thymidine incorporation, a viability/proliferation assay based on metabolic conversion of tetrazolium salt to formazan dye and cell counting. Nicotine administration (10^−6^ M) stimulated cell cycle entry marked by increased DNA synthesis, PCNA and cyclin D1 production, and increased cell division. Nicotinic receptor blockade with d‐tubocurarine, a nicotinic AchR blocker, decreased nicotine‐induced DNA synthesis, and cell division (0.33 ± 0.04, 0.77 ± 0.31‐fold decrease, respectively). Nicotine increased cellular PDGF‐BB transcript levels and protein release (ELISA: 1.6 ± 0.5‐fold increase) but not PDGF‐AA or PDGF–AB release. Nicotine increased PDGFβ‐receptor protein content. PDGF inactivation with anti‐PDGF antibody abolished nicotine‐induced DNA synthesis (1.9 ± 0.08‐fold decrease). PDGF‐R blockade with the PDGF‐R antagonist tyrphostin AG 1295 decreased nicotine‐induced DNA synthesis and cell division (0.25 ± 0.01, 0.44 ± 0.2‐fold decrease, respectively). PDGF‐R blockade reversed nicotine‐stimulated increases in PDGF release, PDGF‐BB transcripts, and PDGF‐receptor levels (0.68 ± 0.34; 0.46 ± 0.01; 0.28 ± 0.01‐fold decrease, respectively). In conclusion, nicotine‐mediated activation of nAChRs increases PDGF‐BB transcription and protein production as well as PDGF β‐receptor levels. PDGF‐BB/PDGF‐R interaction is vital in nicotine's mitogenic actions on human aortic smooth muscle cells. J. Cell. Biochem. © 2005 Wiley‐Liss, Inc.