Nitric oxide inhibits endothelin-1-induced cardiomyocyte hypertrophy through cGMP-mediated suppression of extracellular-signal regulated kinase phosphorylation

Tzu Hurng Cheng, Neng Lang Shih, Shin Yee Chen, Jia Wei Lin, Yen Ling Chen, Cheng Hsien Chen, Heng Lin, Ching Feng Cheng, Wen Ta Chiu, Danny Ling Wang, Jin Jer Chen

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Cardiac hypertrophy is a compensatory mechanism in response to a variety of cardiovascular diseases. Recently, reactive oxygen species and nitric oxide (NO) have been demonstrated to be involved in the pathogenesis of atherosclerosis; however, the role of these free radicals in the development of cardiac hypertrophy remains unclear. In this study, we investigate NO modulation of cellular signaling in endothelin-1 (ET-1)-induced cardiomyocyte hypertrophy in culture. ET-1 treatment of cardiomyocytes increased constitutive NO synthase activity and induced NO production via the stimulation of ET-receptor subtype ET B. Using Northern blot analysis and chloramphenicol acetyltransferase assay, we found that NO suppressed the ET-1-induced increase in c-fos mRNA level and promoter activity. In contrast, ET-1 stimulation of c-fos expression was augmented by depletion of endogenous NO generation with the addition of NO scavenger PTIO into cardiomyocytes. Cells cotransfected with the dominant negative and positive mutants of signaling molecules revealed that the Ras/Raf/extracellularsignal regulated kinase (ERK) signaling pathway is involved in ET-induced c-fos gene expression. Furthermore, NO directly inhibited ET-1-induced ERK phosphorylation and activation in a cGMP-dependent manner, indicating that NO modulates ET-1-induced c-fos expression via its inhibitory effect on ERK signaling pathway. The ET-1-stimulated activator protein-1 (AP-1) DNA binding activity and AP-1-mediated reporter activity were attenuated by NO. In addition, NO also significantly inhibited ET-1-stimulated promoter activity of hypertrophic marker gene β-myosin heavy chain and the enhanced protein synthesis. Taken together, our findings provide the molecular basis of NO as a negative regulator in ET-1-induced cardiac hypertrophy.

Original languageEnglish
Pages (from-to)1183-1192
Number of pages10
JournalMolecular Pharmacology
Volume68
Issue number4
DOIs
Publication statusPublished - Oct 2005
Externally publishedYes

Fingerprint

Extracellular Signal-Regulated MAP Kinases
Endothelin-1
Cardiac Myocytes
Hypertrophy
Nitric Oxide
Phosphorylation
Cardiomegaly
Phosphotransferases
Transcription Factor AP-1
fos Genes
Chloramphenicol O-Acetyltransferase
Myosin Heavy Chains
Nitric Oxide Synthase
Northern Blotting
Free Radicals
Reactive Oxygen Species
Atherosclerosis
Cardiovascular Diseases
Gene Expression
Messenger RNA

ASJC Scopus subject areas

  • Pharmacology

Cite this

Nitric oxide inhibits endothelin-1-induced cardiomyocyte hypertrophy through cGMP-mediated suppression of extracellular-signal regulated kinase phosphorylation. / Cheng, Tzu Hurng; Shih, Neng Lang; Chen, Shin Yee; Lin, Jia Wei; Chen, Yen Ling; Chen, Cheng Hsien; Lin, Heng; Cheng, Ching Feng; Chiu, Wen Ta; Wang, Danny Ling; Chen, Jin Jer.

In: Molecular Pharmacology, Vol. 68, No. 4, 10.2005, p. 1183-1192.

Research output: Contribution to journalArticle

Cheng, Tzu Hurng ; Shih, Neng Lang ; Chen, Shin Yee ; Lin, Jia Wei ; Chen, Yen Ling ; Chen, Cheng Hsien ; Lin, Heng ; Cheng, Ching Feng ; Chiu, Wen Ta ; Wang, Danny Ling ; Chen, Jin Jer. / Nitric oxide inhibits endothelin-1-induced cardiomyocyte hypertrophy through cGMP-mediated suppression of extracellular-signal regulated kinase phosphorylation. In: Molecular Pharmacology. 2005 ; Vol. 68, No. 4. pp. 1183-1192.
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T1 - Nitric oxide inhibits endothelin-1-induced cardiomyocyte hypertrophy through cGMP-mediated suppression of extracellular-signal regulated kinase phosphorylation

AU - Cheng, Tzu Hurng

AU - Shih, Neng Lang

AU - Chen, Shin Yee

AU - Lin, Jia Wei

AU - Chen, Yen Ling

AU - Chen, Cheng Hsien

AU - Lin, Heng

AU - Cheng, Ching Feng

AU - Chiu, Wen Ta

AU - Wang, Danny Ling

AU - Chen, Jin Jer

PY - 2005/10

Y1 - 2005/10

N2 - Cardiac hypertrophy is a compensatory mechanism in response to a variety of cardiovascular diseases. Recently, reactive oxygen species and nitric oxide (NO) have been demonstrated to be involved in the pathogenesis of atherosclerosis; however, the role of these free radicals in the development of cardiac hypertrophy remains unclear. In this study, we investigate NO modulation of cellular signaling in endothelin-1 (ET-1)-induced cardiomyocyte hypertrophy in culture. ET-1 treatment of cardiomyocytes increased constitutive NO synthase activity and induced NO production via the stimulation of ET-receptor subtype ET B. Using Northern blot analysis and chloramphenicol acetyltransferase assay, we found that NO suppressed the ET-1-induced increase in c-fos mRNA level and promoter activity. In contrast, ET-1 stimulation of c-fos expression was augmented by depletion of endogenous NO generation with the addition of NO scavenger PTIO into cardiomyocytes. Cells cotransfected with the dominant negative and positive mutants of signaling molecules revealed that the Ras/Raf/extracellularsignal regulated kinase (ERK) signaling pathway is involved in ET-induced c-fos gene expression. Furthermore, NO directly inhibited ET-1-induced ERK phosphorylation and activation in a cGMP-dependent manner, indicating that NO modulates ET-1-induced c-fos expression via its inhibitory effect on ERK signaling pathway. The ET-1-stimulated activator protein-1 (AP-1) DNA binding activity and AP-1-mediated reporter activity were attenuated by NO. In addition, NO also significantly inhibited ET-1-stimulated promoter activity of hypertrophic marker gene β-myosin heavy chain and the enhanced protein synthesis. Taken together, our findings provide the molecular basis of NO as a negative regulator in ET-1-induced cardiac hypertrophy.

AB - Cardiac hypertrophy is a compensatory mechanism in response to a variety of cardiovascular diseases. Recently, reactive oxygen species and nitric oxide (NO) have been demonstrated to be involved in the pathogenesis of atherosclerosis; however, the role of these free radicals in the development of cardiac hypertrophy remains unclear. In this study, we investigate NO modulation of cellular signaling in endothelin-1 (ET-1)-induced cardiomyocyte hypertrophy in culture. ET-1 treatment of cardiomyocytes increased constitutive NO synthase activity and induced NO production via the stimulation of ET-receptor subtype ET B. Using Northern blot analysis and chloramphenicol acetyltransferase assay, we found that NO suppressed the ET-1-induced increase in c-fos mRNA level and promoter activity. In contrast, ET-1 stimulation of c-fos expression was augmented by depletion of endogenous NO generation with the addition of NO scavenger PTIO into cardiomyocytes. Cells cotransfected with the dominant negative and positive mutants of signaling molecules revealed that the Ras/Raf/extracellularsignal regulated kinase (ERK) signaling pathway is involved in ET-induced c-fos gene expression. Furthermore, NO directly inhibited ET-1-induced ERK phosphorylation and activation in a cGMP-dependent manner, indicating that NO modulates ET-1-induced c-fos expression via its inhibitory effect on ERK signaling pathway. The ET-1-stimulated activator protein-1 (AP-1) DNA binding activity and AP-1-mediated reporter activity were attenuated by NO. In addition, NO also significantly inhibited ET-1-stimulated promoter activity of hypertrophic marker gene β-myosin heavy chain and the enhanced protein synthesis. Taken together, our findings provide the molecular basis of NO as a negative regulator in ET-1-induced cardiac hypertrophy.

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