Gene transfer of nitric oxide synthase: Effects on endothelial biology

Josef Niebauer, Józef Dulak, Jason R. Chan, Philip S. Tsao, John P. Cooke

Research output: Contribution to journalArticlepeer-review

43 Scopus citations


OBJECTIVES: The purpose of the study was to investigate the role of nitric oxide (NO) in monocyte-endothelial interaction by augmenting NO release via transfection of human endothelial cells (ECs) with EC NO synthase (eNOS) DNA. BACKGROUND: Enhancement of NO synthesis by L-arginine or shear stress reduces endothelial adhesiveness for monocytes and inhibits atherogenesis. To elucidate further the underlying mechanism, we augmented NO synthase expression by transfection of human EC. METHODS: Liposome-mediated transfection of EC was performed with a plasmid construct containing the gene encoding eNOS. Expression of eNOS was confirmed by reverse transcription polymerase chain reaction (RT-PCR). Endothelial cells were exposed to human monocytoid cells, and adherent cells were quantitated using a computer- assisted program. Nitric oxide was measured by chemiluminescence. RESULTS: The NO levels were not different in EC that were either not transfected, transfected with beta-gal or liposomes only. The nitric oxide synthase (NOS) transfection increased NO release by +60% (n = 6), which increased further when EC were stimulated by shear stress (24 h) by +137% (n = 5) as compared with untransfected, unstimulated EC (both p < 0.05). The RT-PCR revealed diminished monocyte chemotactic protein-1 (MCP-1) expression in eNOS transfected EC. There was an inverse relation between NO levels and monocyte binding (r = -0.5669, p < 0.002). Stimulation of EC with tumor necrosis factor-alpha (TNF-alpha; 250 U/ml) led to a decrease in NO synthesis, and an increase in monocyte binding. Cells transfected with NOS were resistant to both effects of TNF-alpha. CONCLUSIONS: Endothelial cells transfected with eNOS synthesize an increased amount of NO; this is associated with diminished MCP-1 expression and monocyte-endothelial binding. The reduction in monocyte- endothelial binding persists even after cytokine stimulation.

Original languageEnglish (US)
Pages (from-to)1201-1207
Number of pages7
JournalJournal of the American College of Cardiology
Issue number4
StatePublished - Oct 1999

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine


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