Increased expression of the cGMP-inhibited cAMP-specific (PDE3) and cGMP binding cGMP-specific (PDE5) phosphodiesterases in models of pulmonary hypertension

Fiona Murray, Margaret R MacLean, Nigel J Pyne

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

1. Chronic hypoxic treatment of rats (to induce pulmonary hypertension, PHT) for 14 days increased cGMP-inhibited cAMP specific phosphodiesterase (PDE3) and cGMP binding cGMP specific phosphodiesterase (PDE5) activities in pulmonary arteries. The objective of this study was to establish the molecular basis for these changes in both animal and cell models of PHT. In this regard, RT-PCR and quantitative Western blotting analysis was applied to rat pulmonary artery homogenates and human pulmonary "artery" smooth muscle cell (HPASMC) lysates. 2. PDE3A/B gene transcript levels were increased in the main, first, intrapulmonary and resistance pulmonary arteries by chronic hypoxia. mRNA transcript and protein levels of PDE5A2 in the main and first branch pulmonary arteries were also increased by chronic hypoxia, with no effect on PDE5A1/A2 in the intra-pulmonary and resistance vessels. 3. The expression of PDE3A was increased in HPASMCs maintained under chronic hypoxic conditions for 14 days. This may be mediated via a protein kinase A-dependent mechanism, as treatment of cells with Br-cAMP (100 microM) mimicked chronic hypoxia in increasing PDE3A expression, while the PKA inhibitor, H8 peptide (50 microM) abolished the hypoxic-dependent increase in PDE3A transcript. 4. We also found that the treatment of HPASMCs with the inhibitor of kappaB degradation Tosyl-Leucyl-Chloro-Ketone (TLCK, 50 microM) reduced PDE5 transcript levels, suggesting a role for this transcription factor in the regulation of PDE5 gene expression. 5. Our results show that increased expression of PDE3 and PDE5 might explain some changes in vascular reactivity of pulmonary vessels from rats with PHT. We also report that NF-kappaB might regulate basal PDE5 expression.
LanguageEnglish
Pages1187-1194
Number of pages8
JournalBritish Journal of Pharmacology
Volume137
Issue number8
DOIs
Publication statusPublished - Dec 2002

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Type 5 Cyclic Nucleotide Phosphodiesterases
Pulmonary Hypertension
Pulmonary Artery
Tosyllysine Chloromethyl Ketone
Lung
NF-kappa B
Phosphoric Diester Hydrolases
Gene Expression Regulation
Cyclic AMP-Dependent Protein Kinases
Ketones
varespladib methyl
Smooth Muscle Myocytes
Blood Vessels
Transcription Factors
Animal Models
Western Blotting
Polymerase Chain Reaction
Messenger RNA
Peptides
Genes

Keywords

  • 3',5'-Cyclic-AMP Phosphodiesterases
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • animals
  • cell line
  • cyclic AMP
  • cyclic GMP
  • cyclic nucleotide phosphodiesterases, type 3
  • cyclic nucleotide phosphodiesterases, type 5
  • humans
  • hypertension, pulmonary
  • male
  • myocytes, smooth muscle
  • pulmonary artery
  • rats
  • rats, wistar
  • tosyllysine chloromethyl ketone
  • genes

Cite this

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title = "Increased expression of the cGMP-inhibited cAMP-specific (PDE3) and cGMP binding cGMP-specific (PDE5) phosphodiesterases in models of pulmonary hypertension",
abstract = "1. Chronic hypoxic treatment of rats (to induce pulmonary hypertension, PHT) for 14 days increased cGMP-inhibited cAMP specific phosphodiesterase (PDE3) and cGMP binding cGMP specific phosphodiesterase (PDE5) activities in pulmonary arteries. The objective of this study was to establish the molecular basis for these changes in both animal and cell models of PHT. In this regard, RT-PCR and quantitative Western blotting analysis was applied to rat pulmonary artery homogenates and human pulmonary {"}artery{"} smooth muscle cell (HPASMC) lysates. 2. PDE3A/B gene transcript levels were increased in the main, first, intrapulmonary and resistance pulmonary arteries by chronic hypoxia. mRNA transcript and protein levels of PDE5A2 in the main and first branch pulmonary arteries were also increased by chronic hypoxia, with no effect on PDE5A1/A2 in the intra-pulmonary and resistance vessels. 3. The expression of PDE3A was increased in HPASMCs maintained under chronic hypoxic conditions for 14 days. This may be mediated via a protein kinase A-dependent mechanism, as treatment of cells with Br-cAMP (100 microM) mimicked chronic hypoxia in increasing PDE3A expression, while the PKA inhibitor, H8 peptide (50 microM) abolished the hypoxic-dependent increase in PDE3A transcript. 4. We also found that the treatment of HPASMCs with the inhibitor of kappaB degradation Tosyl-Leucyl-Chloro-Ketone (TLCK, 50 microM) reduced PDE5 transcript levels, suggesting a role for this transcription factor in the regulation of PDE5 gene expression. 5. Our results show that increased expression of PDE3 and PDE5 might explain some changes in vascular reactivity of pulmonary vessels from rats with PHT. We also report that NF-kappaB might regulate basal PDE5 expression.",
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author = "Fiona Murray and MacLean, {Margaret R} and Pyne, {Nigel J}",
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Increased expression of the cGMP-inhibited cAMP-specific (PDE3) and cGMP binding cGMP-specific (PDE5) phosphodiesterases in models of pulmonary hypertension. / Murray, Fiona; MacLean, Margaret R; Pyne, Nigel J.

In: British Journal of Pharmacology, Vol. 137, No. 8, 12.2002, p. 1187-1194.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Increased expression of the cGMP-inhibited cAMP-specific (PDE3) and cGMP binding cGMP-specific (PDE5) phosphodiesterases in models of pulmonary hypertension

AU - Murray, Fiona

AU - MacLean, Margaret R

AU - Pyne, Nigel J

PY - 2002/12

Y1 - 2002/12

N2 - 1. Chronic hypoxic treatment of rats (to induce pulmonary hypertension, PHT) for 14 days increased cGMP-inhibited cAMP specific phosphodiesterase (PDE3) and cGMP binding cGMP specific phosphodiesterase (PDE5) activities in pulmonary arteries. The objective of this study was to establish the molecular basis for these changes in both animal and cell models of PHT. In this regard, RT-PCR and quantitative Western blotting analysis was applied to rat pulmonary artery homogenates and human pulmonary "artery" smooth muscle cell (HPASMC) lysates. 2. PDE3A/B gene transcript levels were increased in the main, first, intrapulmonary and resistance pulmonary arteries by chronic hypoxia. mRNA transcript and protein levels of PDE5A2 in the main and first branch pulmonary arteries were also increased by chronic hypoxia, with no effect on PDE5A1/A2 in the intra-pulmonary and resistance vessels. 3. The expression of PDE3A was increased in HPASMCs maintained under chronic hypoxic conditions for 14 days. This may be mediated via a protein kinase A-dependent mechanism, as treatment of cells with Br-cAMP (100 microM) mimicked chronic hypoxia in increasing PDE3A expression, while the PKA inhibitor, H8 peptide (50 microM) abolished the hypoxic-dependent increase in PDE3A transcript. 4. We also found that the treatment of HPASMCs with the inhibitor of kappaB degradation Tosyl-Leucyl-Chloro-Ketone (TLCK, 50 microM) reduced PDE5 transcript levels, suggesting a role for this transcription factor in the regulation of PDE5 gene expression. 5. Our results show that increased expression of PDE3 and PDE5 might explain some changes in vascular reactivity of pulmonary vessels from rats with PHT. We also report that NF-kappaB might regulate basal PDE5 expression.

AB - 1. Chronic hypoxic treatment of rats (to induce pulmonary hypertension, PHT) for 14 days increased cGMP-inhibited cAMP specific phosphodiesterase (PDE3) and cGMP binding cGMP specific phosphodiesterase (PDE5) activities in pulmonary arteries. The objective of this study was to establish the molecular basis for these changes in both animal and cell models of PHT. In this regard, RT-PCR and quantitative Western blotting analysis was applied to rat pulmonary artery homogenates and human pulmonary "artery" smooth muscle cell (HPASMC) lysates. 2. PDE3A/B gene transcript levels were increased in the main, first, intrapulmonary and resistance pulmonary arteries by chronic hypoxia. mRNA transcript and protein levels of PDE5A2 in the main and first branch pulmonary arteries were also increased by chronic hypoxia, with no effect on PDE5A1/A2 in the intra-pulmonary and resistance vessels. 3. The expression of PDE3A was increased in HPASMCs maintained under chronic hypoxic conditions for 14 days. This may be mediated via a protein kinase A-dependent mechanism, as treatment of cells with Br-cAMP (100 microM) mimicked chronic hypoxia in increasing PDE3A expression, while the PKA inhibitor, H8 peptide (50 microM) abolished the hypoxic-dependent increase in PDE3A transcript. 4. We also found that the treatment of HPASMCs with the inhibitor of kappaB degradation Tosyl-Leucyl-Chloro-Ketone (TLCK, 50 microM) reduced PDE5 transcript levels, suggesting a role for this transcription factor in the regulation of PDE5 gene expression. 5. Our results show that increased expression of PDE3 and PDE5 might explain some changes in vascular reactivity of pulmonary vessels from rats with PHT. We also report that NF-kappaB might regulate basal PDE5 expression.

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KW - 3',5'-Cyclic-GMP Phosphodiesterases

KW - animals

KW - cell line

KW - cyclic AMP

KW - cyclic GMP

KW - cyclic nucleotide phosphodiesterases, type 3

KW - cyclic nucleotide phosphodiesterases, type 5

KW - humans

KW - hypertension, pulmonary

KW - male

KW - myocytes, smooth muscle

KW - pulmonary artery

KW - rats

KW - rats, wistar

KW - tosyllysine chloromethyl ketone

KW - genes

U2 - 10.1038/sj.bjp.0704984

DO - 10.1038/sj.bjp.0704984

M3 - Article

VL - 137

SP - 1187

EP - 1194

JO - British Journal of Pharmacology

T2 - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 8

ER -