Enhanced endothelium-dependent relaxations after gene transfer of recombinant endothelial nitric oxide synthase to rabbit carotid arteries

Iftikhar Jan Kullo, Geza Mozes, Robert S. Schwartz, Peter Gloviczki, Masato Tsutsui, Zvonimir S Katusic, Timothy O'Brien

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

We tested the effects of overexpression of the endothelial nitric oxide synthase (eNOS) gene in the normal arterial wall by adenoviral-mediated gene transfer. Rabbit carotid arteries were surgically isolated and exposed to adenoviral vectors encoding eNOS (AdeNOS) or β-galactosidase (AdβGal) on the contralateral side. Vector solutions at a concentration of 1 x 1010 plaque forming units/mL were instilled for 20 minutes before restoration of flow. Arteries were harvested 4 days later for immunostaining, measurement of cGMP, and vasomotor studies. Endothelium-specific gene transfer was confirmed by staining for β-galactosidase in the AdβGal arteries. Immunostaining of en face endothelial cell imprints from AdeNOS-transduced arteries with a monoclonal antibody to eNOS showed increased immunoreactivity. Basal cGMp levels were significantly greater in the AdeNOS-transduced arteries (18.4±4.6 versus 4.2±0.5 pmol/mg protein; P<.05). Contractions to phenylephrine were significantly reduced in the AdeNOS-transduced arteries (area under curve, 106±5 versus 119±7; P<.05), but in the presence of the eNOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA, 3x10-4 mol/L), there was no difference between the two (area under curve, 148±5 versus 153±6; P=NS). Relaxations to acetylcholine obtained during submaximal contractions to phenylephrine were significantly enhanced in the AdeNOS-transduced arteries (EC50, 7.45±0.05 versus 7.23±0.03; P<.05). We conclude that overexpression of eNOS in the endothelium results in diminished contractile responses, as well as enhanced endothelium-dependent relaxations. These findings imply a possible role for vascular eNOS gene transfer in the treatment of vasospasm and endothelial dysfunction.

Original languageEnglish (US)
Pages (from-to)314-320
Number of pages7
JournalHypertension
Volume30
Issue number3 I
StatePublished - Sep 1997

Fingerprint

Nitric Oxide Synthase Type III
Carotid Arteries
Endothelium
Arteries
Galactosidases
Rabbits
Genes
Phenylephrine
Area Under Curve
omega-N-Methylarginine
Acetylcholine
Blood Vessels
Arginine
Endothelial Cells
Monoclonal Antibodies
Staining and Labeling
Proteins

Keywords

  • Adenovirus
  • Endothelium
  • Gene transfer
  • Nitric oxide
  • Nitric oxide synthase

ASJC Scopus subject areas

  • Internal Medicine

Cite this

Enhanced endothelium-dependent relaxations after gene transfer of recombinant endothelial nitric oxide synthase to rabbit carotid arteries. / Kullo, Iftikhar Jan; Mozes, Geza; Schwartz, Robert S.; Gloviczki, Peter; Tsutsui, Masato; Katusic, Zvonimir S; O'Brien, Timothy.

In: Hypertension, Vol. 30, No. 3 I, 09.1997, p. 314-320.

Research output: Contribution to journalArticle

Kullo, Iftikhar Jan ; Mozes, Geza ; Schwartz, Robert S. ; Gloviczki, Peter ; Tsutsui, Masato ; Katusic, Zvonimir S ; O'Brien, Timothy. / Enhanced endothelium-dependent relaxations after gene transfer of recombinant endothelial nitric oxide synthase to rabbit carotid arteries. In: Hypertension. 1997 ; Vol. 30, No. 3 I. pp. 314-320.
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T1 - Enhanced endothelium-dependent relaxations after gene transfer of recombinant endothelial nitric oxide synthase to rabbit carotid arteries

AU - Kullo, Iftikhar Jan

AU - Mozes, Geza

AU - Schwartz, Robert S.

AU - Gloviczki, Peter

AU - Tsutsui, Masato

AU - Katusic, Zvonimir S

AU - O'Brien, Timothy

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AB - We tested the effects of overexpression of the endothelial nitric oxide synthase (eNOS) gene in the normal arterial wall by adenoviral-mediated gene transfer. Rabbit carotid arteries were surgically isolated and exposed to adenoviral vectors encoding eNOS (AdeNOS) or β-galactosidase (AdβGal) on the contralateral side. Vector solutions at a concentration of 1 x 1010 plaque forming units/mL were instilled for 20 minutes before restoration of flow. Arteries were harvested 4 days later for immunostaining, measurement of cGMP, and vasomotor studies. Endothelium-specific gene transfer was confirmed by staining for β-galactosidase in the AdβGal arteries. Immunostaining of en face endothelial cell imprints from AdeNOS-transduced arteries with a monoclonal antibody to eNOS showed increased immunoreactivity. Basal cGMp levels were significantly greater in the AdeNOS-transduced arteries (18.4±4.6 versus 4.2±0.5 pmol/mg protein; P<.05). Contractions to phenylephrine were significantly reduced in the AdeNOS-transduced arteries (area under curve, 106±5 versus 119±7; P<.05), but in the presence of the eNOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA, 3x10-4 mol/L), there was no difference between the two (area under curve, 148±5 versus 153±6; P=NS). Relaxations to acetylcholine obtained during submaximal contractions to phenylephrine were significantly enhanced in the AdeNOS-transduced arteries (EC50, 7.45±0.05 versus 7.23±0.03; P<.05). We conclude that overexpression of eNOS in the endothelium results in diminished contractile responses, as well as enhanced endothelium-dependent relaxations. These findings imply a possible role for vascular eNOS gene transfer in the treatment of vasospasm and endothelial dysfunction.

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