Expression and function of recombinant endothelial nitric oxide synthase in human endothelial cells

Michela Zanetti, Zvonimir S Katusic, Timothy O'Brien

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Endothelial dysfunction is frequently involved in the pathogenesis of vascular disease. While nitric oxide (NO) inhibits smooth muscle cell proliferation, its effect on endothelial cell proliferation is unclear. The aim of this study was to determine if adenoviral-mediated gene transfer of endothelial NO synthase (eNOS) to human umbilical vein endothelial cells (HUVECs) would result in increased generation of NO and affect endothelial cell proliferation. HUVECs were transduced with adenoviral vectors encoding eNOS (AdeNOS) or β-galactosidase (Adβgal) or exposed to diluent (control). AdeNOS-transduced cells showed increased eNOS expression as detected by Western blot analysis, and increased concentrations of cGMP (control 0.7 ± 0.1; Adβgal 0.9 ± 0.2; AdeNOS 3.1 ± 0.5 pmol/mg protein; p < 0.001) and nitrite (control 11.8 ± 1.2; Adβgal 13.3 ± 1.7; AdeNOS 21.1 ± 2.2 nmol/mg protein/hour; p<0.01). DNA synthesis as assessed by [3H]thymidine incorporation and cell counts were significantly reduced (by ∼30%) in AdeNOS-transduced HUVECs. Expression of mitogen-activated protein kinase was also decreased in AdeNOS-transduced cells. This study shows that adenoviral-mediated gene transfer of eNOS to HUVECs inhibits endothelial cell proliferation.

Original languageEnglish (US)
Pages (from-to)449-456
Number of pages8
JournalJournal of Vascular Research
Volume37
Issue number6
DOIs
StatePublished - 2000

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Nitric Oxide Synthase Type III
Human Umbilical Vein Endothelial Cells
Galactosidases
Endothelial Cells
Cell Proliferation
Nitric Oxide Synthase
Nitric Oxide
Nitrites
Mitogen-Activated Protein Kinases
Vascular Diseases
Thymidine
Genes
Smooth Muscle Myocytes
Proteins
Cell Count
Western Blotting
DNA

Keywords

  • Cell culture/isolation
  • Endothelial factors
  • Endothelial function
  • Gene therapy
  • Nitric oxide

ASJC Scopus subject areas

  • Physiology

Cite this

Expression and function of recombinant endothelial nitric oxide synthase in human endothelial cells. / Zanetti, Michela; Katusic, Zvonimir S; O'Brien, Timothy.

In: Journal of Vascular Research, Vol. 37, No. 6, 2000, p. 449-456.

Research output: Contribution to journalArticle

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