Downregulation of cGMP-dependent protein kinase expression by inflammatory cytokines in vascular smooth muscle cells

Natasha C. Browner, Hassan Sellak, Thomas M. Lincoln

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

NO and cGMP have antigrowth and anti-inflammatory effects on the vessel wall in response to injury. It is well established that after vascular injury proinflammatory cytokines are involved in vascular wall remodeling. The purpose of this study was to ascertain the signaling mechanisms involved in cGMP-dependent protein kinase (PKG) suppression by inflammatory cytokines in primary bovine aortic vascular smooth muscle cells (VSMC). Interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and LPS decreased the mRNA and protein levels of PKG in VSMC. IL-1β, TNF-α, and LPS increased inducible nitric oxide synthase (iNOS) expression and cGMP production. Treatment of cells with selective inhibitors of iNOS or soluble guanylate cyclase (sGC) reversed the downregulation of PKG expression induced by cytokines and LPS. The NO donor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2- diolate (DETA NONOate) and 3-(5-hydroxymethyl-2-furyl)-1-benzylindazole (YC-1), a NO-independent sGC activator, decreased PKG mRNA and protein expression in bovine aortic VSMC. Cyclic nucleotide analogs [8-(4-chlorophenylthio)guanosine 3′,5′-cyclic monophosphate (CPT-cGMP) and 8-(4-chlorophenylthio) adenosine 3,5′-cyclic monophosphate (CPT-cAMP)] also suppressed PKG mRNA and protein expression. However, CPT-cAMP was more effective than CPT-cGMP in decreasing PKG mRNA levels. Selective inhibition of PKA with the Rp isomer of 8-(4-chlorophenylthio)-adenosine 3′,5′-cyclic monophosphorothioate (Rp-8p-CPT cAMPS) prevented the downregulation of PKG by LPS. In contrast, the Rp isomer of 8-(4-chlorophenylthio)guanosine 3,5′-cyclic monophosphorothioate (Rp-8p-CPT cGMPS; inhibitor of PKG) had no effect on LPS-induced inhibition of PKG mRNA and protein expression. These studies suggest that cross-activation of PKA in response to iNOS expression by inflammatory mediators downregulates PKG expression in bovine aortic VSMC.

Original languageEnglish (US)
Pages (from-to)C88-C96
JournalAmerican Journal of Physiology - Cell Physiology
Volume287
Issue number1 56-1
DOIs
StatePublished - Jul 1 2004

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Cyclic GMP-Dependent Protein Kinases
Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Down-Regulation
Cells
Cytokines
Nitric Oxide Synthase Type II
Messenger RNA
Guanosine
Guanylate Cyclase
Interleukin-1
Isomers
Adenosine
Proteins
Tumor Necrosis Factor-alpha
Vascular System Injuries
Cyclic Nucleotides
Anti-Inflammatory Agents
Chemical activation

Keywords

  • Inflammation
  • Nitric oxide
  • Vascular injury

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

Cite this

Downregulation of cGMP-dependent protein kinase expression by inflammatory cytokines in vascular smooth muscle cells. / Browner, Natasha C.; Sellak, Hassan; Lincoln, Thomas M.

In: American Journal of Physiology - Cell Physiology, Vol. 287, No. 1 56-1, 01.07.2004, p. C88-C96.

Research output: Contribution to journalArticle

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N2 - NO and cGMP have antigrowth and anti-inflammatory effects on the vessel wall in response to injury. It is well established that after vascular injury proinflammatory cytokines are involved in vascular wall remodeling. The purpose of this study was to ascertain the signaling mechanisms involved in cGMP-dependent protein kinase (PKG) suppression by inflammatory cytokines in primary bovine aortic vascular smooth muscle cells (VSMC). Interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and LPS decreased the mRNA and protein levels of PKG in VSMC. IL-1β, TNF-α, and LPS increased inducible nitric oxide synthase (iNOS) expression and cGMP production. Treatment of cells with selective inhibitors of iNOS or soluble guanylate cyclase (sGC) reversed the downregulation of PKG expression induced by cytokines and LPS. The NO donor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2- diolate (DETA NONOate) and 3-(5-hydroxymethyl-2-furyl)-1-benzylindazole (YC-1), a NO-independent sGC activator, decreased PKG mRNA and protein expression in bovine aortic VSMC. Cyclic nucleotide analogs [8-(4-chlorophenylthio)guanosine 3′,5′-cyclic monophosphate (CPT-cGMP) and 8-(4-chlorophenylthio) adenosine 3,5′-cyclic monophosphate (CPT-cAMP)] also suppressed PKG mRNA and protein expression. However, CPT-cAMP was more effective than CPT-cGMP in decreasing PKG mRNA levels. Selective inhibition of PKA with the Rp isomer of 8-(4-chlorophenylthio)-adenosine 3′,5′-cyclic monophosphorothioate (Rp-8p-CPT cAMPS) prevented the downregulation of PKG by LPS. In contrast, the Rp isomer of 8-(4-chlorophenylthio)guanosine 3,5′-cyclic monophosphorothioate (Rp-8p-CPT cGMPS; inhibitor of PKG) had no effect on LPS-induced inhibition of PKG mRNA and protein expression. These studies suggest that cross-activation of PKA in response to iNOS expression by inflammatory mediators downregulates PKG expression in bovine aortic VSMC.

AB - NO and cGMP have antigrowth and anti-inflammatory effects on the vessel wall in response to injury. It is well established that after vascular injury proinflammatory cytokines are involved in vascular wall remodeling. The purpose of this study was to ascertain the signaling mechanisms involved in cGMP-dependent protein kinase (PKG) suppression by inflammatory cytokines in primary bovine aortic vascular smooth muscle cells (VSMC). Interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and LPS decreased the mRNA and protein levels of PKG in VSMC. IL-1β, TNF-α, and LPS increased inducible nitric oxide synthase (iNOS) expression and cGMP production. Treatment of cells with selective inhibitors of iNOS or soluble guanylate cyclase (sGC) reversed the downregulation of PKG expression induced by cytokines and LPS. The NO donor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2- diolate (DETA NONOate) and 3-(5-hydroxymethyl-2-furyl)-1-benzylindazole (YC-1), a NO-independent sGC activator, decreased PKG mRNA and protein expression in bovine aortic VSMC. Cyclic nucleotide analogs [8-(4-chlorophenylthio)guanosine 3′,5′-cyclic monophosphate (CPT-cGMP) and 8-(4-chlorophenylthio) adenosine 3,5′-cyclic monophosphate (CPT-cAMP)] also suppressed PKG mRNA and protein expression. However, CPT-cAMP was more effective than CPT-cGMP in decreasing PKG mRNA levels. Selective inhibition of PKA with the Rp isomer of 8-(4-chlorophenylthio)-adenosine 3′,5′-cyclic monophosphorothioate (Rp-8p-CPT cAMPS) prevented the downregulation of PKG by LPS. In contrast, the Rp isomer of 8-(4-chlorophenylthio)guanosine 3,5′-cyclic monophosphorothioate (Rp-8p-CPT cGMPS; inhibitor of PKG) had no effect on LPS-induced inhibition of PKG mRNA and protein expression. These studies suggest that cross-activation of PKA in response to iNOS expression by inflammatory mediators downregulates PKG expression in bovine aortic VSMC.

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