Overexpression of endothelium nitric oxide restores relaxation in the hindlimb vasculature after myocardial infarction

M. A. Gaballa, C. Jacob, Steven Goldman

Research output: Contribution to journalArticle

Abstract

After myocardial infarction (MI), NO-mediated vasorelaxation is attenuated in both large and resistance arteries. The mechanisms of this depression are unclear. To determine if the attenuated vasorelaxation after MI is due to downregulation of eNOS protein, immunoblotting and gene transfer of eNOS was performed using a "first-generation" serotype 5, replication-deficient, adenoviral vector (1.2×109 Pfus) containing cDNA encoding the eNOS gene in the hindlimb vasculature for 30 minutes in rats 3 weeks after MI. Five days after infection, overexpression was confirmed by immunohistochemical staining and immunoblotting. Recombinant gene expression was localized to the vascular endothelial cells. ENOS protein level decreased after MI (3.3±0.9 vs. 2.1±0.8 intensity units/μg protein, n=6, P<0.05) and increased two fold (4.3±1.2 intensity units/μg protein, n=5) after gene transfer. Acetylcholine (ACh)-stimulated vasorelaxation in the hindlimb was decreased (P<0.05) by 30% after MI and was restored to normal in MI rats transfected with eNOS. Addition of 100 μM NG-nitro-L-arginine methyl ester (L-NAME) abolished the difference between sham, MI, and MI transfected rats. The attenuated NO-mediated vasorelaxation in the hindlimb after MI is due to a downregulation of eNOS protein. Overexpression of eNOS restores the ACh-mediated vasorelaxation.

Original languageEnglish (US)
JournalJournal of Investigative Medicine
Volume47
Issue number2
StatePublished - Feb 1999

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Hindlimb
Endothelium
Nitric Oxide
Myocardial Infarction
Vasodilation
Gene transfer
Rats
NG-Nitroarginine Methyl Ester
Acetylcholine
Proteins
Endothelial cells
Immunoblotting
Gene expression
Down-Regulation
Complementary DNA
Genes
Endothelial Cells
Arteries
Staining and Labeling
Gene Expression

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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title = "Overexpression of endothelium nitric oxide restores relaxation in the hindlimb vasculature after myocardial infarction",
abstract = "After myocardial infarction (MI), NO-mediated vasorelaxation is attenuated in both large and resistance arteries. The mechanisms of this depression are unclear. To determine if the attenuated vasorelaxation after MI is due to downregulation of eNOS protein, immunoblotting and gene transfer of eNOS was performed using a {"}first-generation{"} serotype 5, replication-deficient, adenoviral vector (1.2×109 Pfus) containing cDNA encoding the eNOS gene in the hindlimb vasculature for 30 minutes in rats 3 weeks after MI. Five days after infection, overexpression was confirmed by immunohistochemical staining and immunoblotting. Recombinant gene expression was localized to the vascular endothelial cells. ENOS protein level decreased after MI (3.3±0.9 vs. 2.1±0.8 intensity units/μg protein, n=6, P<0.05) and increased two fold (4.3±1.2 intensity units/μg protein, n=5) after gene transfer. Acetylcholine (ACh)-stimulated vasorelaxation in the hindlimb was decreased (P<0.05) by 30{\%} after MI and was restored to normal in MI rats transfected with eNOS. Addition of 100 μM NG-nitro-L-arginine methyl ester (L-NAME) abolished the difference between sham, MI, and MI transfected rats. The attenuated NO-mediated vasorelaxation in the hindlimb after MI is due to a downregulation of eNOS protein. Overexpression of eNOS restores the ACh-mediated vasorelaxation.",
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T1 - Overexpression of endothelium nitric oxide restores relaxation in the hindlimb vasculature after myocardial infarction

AU - Gaballa, M. A.

AU - Jacob, C.

AU - Goldman, Steven

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Y1 - 1999/2

N2 - After myocardial infarction (MI), NO-mediated vasorelaxation is attenuated in both large and resistance arteries. The mechanisms of this depression are unclear. To determine if the attenuated vasorelaxation after MI is due to downregulation of eNOS protein, immunoblotting and gene transfer of eNOS was performed using a "first-generation" serotype 5, replication-deficient, adenoviral vector (1.2×109 Pfus) containing cDNA encoding the eNOS gene in the hindlimb vasculature for 30 minutes in rats 3 weeks after MI. Five days after infection, overexpression was confirmed by immunohistochemical staining and immunoblotting. Recombinant gene expression was localized to the vascular endothelial cells. ENOS protein level decreased after MI (3.3±0.9 vs. 2.1±0.8 intensity units/μg protein, n=6, P<0.05) and increased two fold (4.3±1.2 intensity units/μg protein, n=5) after gene transfer. Acetylcholine (ACh)-stimulated vasorelaxation in the hindlimb was decreased (P<0.05) by 30% after MI and was restored to normal in MI rats transfected with eNOS. Addition of 100 μM NG-nitro-L-arginine methyl ester (L-NAME) abolished the difference between sham, MI, and MI transfected rats. The attenuated NO-mediated vasorelaxation in the hindlimb after MI is due to a downregulation of eNOS protein. Overexpression of eNOS restores the ACh-mediated vasorelaxation.

AB - After myocardial infarction (MI), NO-mediated vasorelaxation is attenuated in both large and resistance arteries. The mechanisms of this depression are unclear. To determine if the attenuated vasorelaxation after MI is due to downregulation of eNOS protein, immunoblotting and gene transfer of eNOS was performed using a "first-generation" serotype 5, replication-deficient, adenoviral vector (1.2×109 Pfus) containing cDNA encoding the eNOS gene in the hindlimb vasculature for 30 minutes in rats 3 weeks after MI. Five days after infection, overexpression was confirmed by immunohistochemical staining and immunoblotting. Recombinant gene expression was localized to the vascular endothelial cells. ENOS protein level decreased after MI (3.3±0.9 vs. 2.1±0.8 intensity units/μg protein, n=6, P<0.05) and increased two fold (4.3±1.2 intensity units/μg protein, n=5) after gene transfer. Acetylcholine (ACh)-stimulated vasorelaxation in the hindlimb was decreased (P<0.05) by 30% after MI and was restored to normal in MI rats transfected with eNOS. Addition of 100 μM NG-nitro-L-arginine methyl ester (L-NAME) abolished the difference between sham, MI, and MI transfected rats. The attenuated NO-mediated vasorelaxation in the hindlimb after MI is due to a downregulation of eNOS protein. Overexpression of eNOS restores the ACh-mediated vasorelaxation.

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