Atrial natriuretic peptide inhibits transforming growth factor β-induced Smad signaling and myofibroblast transformation in mouse cardiac fibroblasts

Peng Li, Dajun Wang, Jason Lucas, Suzanne Oparil, Dongqi Xing, Xu Cao, Lea Novak, Matthew B. Renfrow, Yiu Fai Chen

Research output: Contribution to journalArticle

Abstract

This study tested the hypothesis that activation of atrial natriuretic peptide (ANP)/cGMP/protein kinase G signaling inhibits transforming growth factor (TGF)-β1-induced extracellular matrix expression in cardiac fibroblasts and defined the specific site(s) at which this molecular merging of signaling pathways occurs. Left ventricular hypertrophy and fibrosis, collagen deposition, and myofibroblast transformation of cardiac fibroblasts in response to pressure overload by transverse aortic constriction were exaggerated in ANP-null mice compared with wild-type controls. ANP and cGMP inhibited TGF-β1-induced myofibroblast transformation, proliferation, collagen synthesis, and plasminogen activator inhibitor-1 expression in cardiac fibroblasts isolated from wild-type mice. Following pretreatment with cGMP, TGF-β1 induced phosphorylation of Smad3, but the resultant pSmad3 could not be translocated to the nucleus. pSmad3 that had been phosphorylated with recombinant protein kinase G-1α was analyzed by use of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and ion trap tandem mass spectrometry. The analysis revealed phosphorylation of Ser309 and Thr388 residues, sites distinct from the C-terminal Ser423/425 residues that are phosphorylated by TGF-β receptor kinase and are critical for the nuclear translocation and down-stream signaling of pSmad3. These results suggest that phosphorylation of Smad3 by protein kinase G is a potential molecular mechanism by which activation of ANP/cGMP/protein kinase G signaling disrupts TGF-β1-induced nuclear translocation of pSmad3 and downstream events, including myofibroblast transformation, proliferation, and expression of extracellular matrix molecules in cardiac fibroblasts. We postulate that this process contributes to the antifibrogenic effects of the natriuretic peptide in heart.

Original languageEnglish (US)
Pages (from-to)185-192
Number of pages8
JournalCirculation Research
Volume102
Issue number2
DOIs
StatePublished - Feb 2008
Externally publishedYes

Fingerprint

Myofibroblasts
Transforming Growth Factors
Atrial Natriuretic Factor
Cyclic GMP-Dependent Protein Kinases
Fibroblasts
Phosphorylation
Extracellular Matrix
Smad3 Protein
Collagen
Ions
Cyclotrons
Natriuretic Peptides
Growth Factor Receptors
Plasminogen Activator Inhibitor 1
Left Ventricular Hypertrophy
Fourier Analysis
Tandem Mass Spectrometry
Recombinant Proteins
Constriction
Mass Spectrometry

Keywords

  • Atrial natriuretic factor
  • Cardiac fibroblast
  • Cardiac fibrosis and remodeling
  • Signal transduction
  • Transforming growth factor

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Atrial natriuretic peptide inhibits transforming growth factor β-induced Smad signaling and myofibroblast transformation in mouse cardiac fibroblasts. / Li, Peng; Wang, Dajun; Lucas, Jason; Oparil, Suzanne; Xing, Dongqi; Cao, Xu; Novak, Lea; Renfrow, Matthew B.; Chen, Yiu Fai.

In: Circulation Research, Vol. 102, No. 2, 02.2008, p. 185-192.

Research output: Contribution to journalArticle

Li, Peng ; Wang, Dajun ; Lucas, Jason ; Oparil, Suzanne ; Xing, Dongqi ; Cao, Xu ; Novak, Lea ; Renfrow, Matthew B. ; Chen, Yiu Fai. / Atrial natriuretic peptide inhibits transforming growth factor β-induced Smad signaling and myofibroblast transformation in mouse cardiac fibroblasts. In: Circulation Research. 2008 ; Vol. 102, No. 2. pp. 185-192.
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AB - This study tested the hypothesis that activation of atrial natriuretic peptide (ANP)/cGMP/protein kinase G signaling inhibits transforming growth factor (TGF)-β1-induced extracellular matrix expression in cardiac fibroblasts and defined the specific site(s) at which this molecular merging of signaling pathways occurs. Left ventricular hypertrophy and fibrosis, collagen deposition, and myofibroblast transformation of cardiac fibroblasts in response to pressure overload by transverse aortic constriction were exaggerated in ANP-null mice compared with wild-type controls. ANP and cGMP inhibited TGF-β1-induced myofibroblast transformation, proliferation, collagen synthesis, and plasminogen activator inhibitor-1 expression in cardiac fibroblasts isolated from wild-type mice. Following pretreatment with cGMP, TGF-β1 induced phosphorylation of Smad3, but the resultant pSmad3 could not be translocated to the nucleus. pSmad3 that had been phosphorylated with recombinant protein kinase G-1α was analyzed by use of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and ion trap tandem mass spectrometry. The analysis revealed phosphorylation of Ser309 and Thr388 residues, sites distinct from the C-terminal Ser423/425 residues that are phosphorylated by TGF-β receptor kinase and are critical for the nuclear translocation and down-stream signaling of pSmad3. These results suggest that phosphorylation of Smad3 by protein kinase G is a potential molecular mechanism by which activation of ANP/cGMP/protein kinase G signaling disrupts TGF-β1-induced nuclear translocation of pSmad3 and downstream events, including myofibroblast transformation, proliferation, and expression of extracellular matrix molecules in cardiac fibroblasts. We postulate that this process contributes to the antifibrogenic effects of the natriuretic peptide in heart.

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