Peroxynitrite stimulates pulmonary artery endothelial and smooth muscle cell proliferation: Involvement of ERK and PKC

E. O. Agbani, P. Coats, A. Mills, R. M. Wadsworth

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

There is evidence that peroxynitrite is generated in pulmonary hypertension and we have therefore investigated whether peroxynitrite can cause proliferation of pulmonary artery cells.
Bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC) were exposed to peroxynitrite solution or to the peroxynitrite generating compound, 3-morpholinosydnonimine (SIN-1). Vascular cell proliferation was determined by cell count and H-3-thymidine incorporation. Protein biochemistry was by western blot analysis.
Transient exposure to peroxynitrite stimulated the proliferation of PASMC (peroxynitrite 0.2 nM -2 mu M) and PAEC (peroxynitrite 0.2 mu M). Peroxynitrite 0.2 mu M stimulated DNA synthesis in PASMC cell by 200 +/- 22% and in PAEC by 137 +/- 4%. DNA synthesis in PAEC and PASMC was also stimulated by the peroxynitrite generator SIN-1 2 mu M. Cell proliferation was accompanied by activation of ERK, which peaked at 15 min and remained elevated for 12 h in PASMC. However peroxynitrite at the concentrations used in this study did not activate the stress pathways p38 mitogen activated protein kinase (MAPK) or Jun N-terminal kinase (JNK). Peroxynitrite-induced proliferation and ERK phosphorylation in PASMC were abolished by the peroxynitrite scavenger ebselen 5 mu M. Peroxynitrite-induced proliferation and extracellular signal-regulated kinase (ERK) phosphorylation in PASMC was prevented by selective inhibitors of MAP kinase kinase (MEK) (U0126 5 mu M, PD98059 50 mu M), Raf-1 (Raf-1 kinase inhibitor 10 mu M), Ras (FPT II and FPT III 10 mu M) and protein kinase C (PKC) (GF109203X 10 mu M). Inhibition of EGF or PDGF receptor signaling using AG-1296. AG-1478 or imatinib prevented peroxynitrite-induced cell proliferation and ERK phosphorylation in PASMC.
Peroxynitrite can stimulate proliferation of pulmonary artery cells, involving ERK, PKC and EGF or PDGF receptors.

LanguageEnglish
Pages100-109
Number of pages10
JournalPulmonary Pharmacology and Therapeutics
Volume24
Issue number1
DOIs
Publication statusPublished - Feb 2011

Fingerprint

Peroxynitrous Acid
Extracellular Signal-Regulated MAP Kinases
Cell proliferation
Protein Kinase C
Pulmonary Artery
Smooth Muscle Myocytes
Muscle
Cell Proliferation
Phosphorylation
Platelet-Derived Growth Factor Receptors
MAP Kinase Kinase 5
Epidermal Growth Factor
Proto-Oncogene Proteins c-raf
Biochemistry
DNA
Mitogen-Activated Protein Kinase Kinases
p38 Mitogen-Activated Protein Kinases
Pulmonary Hypertension
Thymidine

Keywords

  • pulmonary
  • hypertension
  • peroxynitrite
  • endothelial cells
  • smooth muscle cells
  • activated protein-kinase
  • signal-transduction
  • oxidative stress
  • nitric-oxide
  • inhibitor
  • receptor
  • pathway
  • RAF-1
  • RAS

Cite this

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abstract = "There is evidence that peroxynitrite is generated in pulmonary hypertension and we have therefore investigated whether peroxynitrite can cause proliferation of pulmonary artery cells. Bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC) were exposed to peroxynitrite solution or to the peroxynitrite generating compound, 3-morpholinosydnonimine (SIN-1). Vascular cell proliferation was determined by cell count and H-3-thymidine incorporation. Protein biochemistry was by western blot analysis. Transient exposure to peroxynitrite stimulated the proliferation of PASMC (peroxynitrite 0.2 nM -2 mu M) and PAEC (peroxynitrite 0.2 mu M). Peroxynitrite 0.2 mu M stimulated DNA synthesis in PASMC cell by 200 +/- 22{\%} and in PAEC by 137 +/- 4{\%}. DNA synthesis in PAEC and PASMC was also stimulated by the peroxynitrite generator SIN-1 2 mu M. Cell proliferation was accompanied by activation of ERK, which peaked at 15 min and remained elevated for 12 h in PASMC. However peroxynitrite at the concentrations used in this study did not activate the stress pathways p38 mitogen activated protein kinase (MAPK) or Jun N-terminal kinase (JNK). Peroxynitrite-induced proliferation and ERK phosphorylation in PASMC were abolished by the peroxynitrite scavenger ebselen 5 mu M. Peroxynitrite-induced proliferation and extracellular signal-regulated kinase (ERK) phosphorylation in PASMC was prevented by selective inhibitors of MAP kinase kinase (MEK) (U0126 5 mu M, PD98059 50 mu M), Raf-1 (Raf-1 kinase inhibitor 10 mu M), Ras (FPT II and FPT III 10 mu M) and protein kinase C (PKC) (GF109203X 10 mu M). Inhibition of EGF or PDGF receptor signaling using AG-1296. AG-1478 or imatinib prevented peroxynitrite-induced cell proliferation and ERK phosphorylation in PASMC. Peroxynitrite can stimulate proliferation of pulmonary artery cells, involving ERK, PKC and EGF or PDGF receptors.",
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Peroxynitrite stimulates pulmonary artery endothelial and smooth muscle cell proliferation : Involvement of ERK and PKC. / Agbani, E. O.; Coats, P.; Mills, A.; Wadsworth, R. M.

In: Pulmonary Pharmacology and Therapeutics, Vol. 24, No. 1, 02.2011, p. 100-109.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Peroxynitrite stimulates pulmonary artery endothelial and smooth muscle cell proliferation

T2 - Pulmonary Pharmacology and Therapeutics

AU - Agbani, E. O.

AU - Coats, P.

AU - Mills, A.

AU - Wadsworth, R. M.

PY - 2011/2

Y1 - 2011/2

N2 - There is evidence that peroxynitrite is generated in pulmonary hypertension and we have therefore investigated whether peroxynitrite can cause proliferation of pulmonary artery cells. Bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC) were exposed to peroxynitrite solution or to the peroxynitrite generating compound, 3-morpholinosydnonimine (SIN-1). Vascular cell proliferation was determined by cell count and H-3-thymidine incorporation. Protein biochemistry was by western blot analysis. Transient exposure to peroxynitrite stimulated the proliferation of PASMC (peroxynitrite 0.2 nM -2 mu M) and PAEC (peroxynitrite 0.2 mu M). Peroxynitrite 0.2 mu M stimulated DNA synthesis in PASMC cell by 200 +/- 22% and in PAEC by 137 +/- 4%. DNA synthesis in PAEC and PASMC was also stimulated by the peroxynitrite generator SIN-1 2 mu M. Cell proliferation was accompanied by activation of ERK, which peaked at 15 min and remained elevated for 12 h in PASMC. However peroxynitrite at the concentrations used in this study did not activate the stress pathways p38 mitogen activated protein kinase (MAPK) or Jun N-terminal kinase (JNK). Peroxynitrite-induced proliferation and ERK phosphorylation in PASMC were abolished by the peroxynitrite scavenger ebselen 5 mu M. Peroxynitrite-induced proliferation and extracellular signal-regulated kinase (ERK) phosphorylation in PASMC was prevented by selective inhibitors of MAP kinase kinase (MEK) (U0126 5 mu M, PD98059 50 mu M), Raf-1 (Raf-1 kinase inhibitor 10 mu M), Ras (FPT II and FPT III 10 mu M) and protein kinase C (PKC) (GF109203X 10 mu M). Inhibition of EGF or PDGF receptor signaling using AG-1296. AG-1478 or imatinib prevented peroxynitrite-induced cell proliferation and ERK phosphorylation in PASMC. Peroxynitrite can stimulate proliferation of pulmonary artery cells, involving ERK, PKC and EGF or PDGF receptors.

AB - There is evidence that peroxynitrite is generated in pulmonary hypertension and we have therefore investigated whether peroxynitrite can cause proliferation of pulmonary artery cells. Bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC) were exposed to peroxynitrite solution or to the peroxynitrite generating compound, 3-morpholinosydnonimine (SIN-1). Vascular cell proliferation was determined by cell count and H-3-thymidine incorporation. Protein biochemistry was by western blot analysis. Transient exposure to peroxynitrite stimulated the proliferation of PASMC (peroxynitrite 0.2 nM -2 mu M) and PAEC (peroxynitrite 0.2 mu M). Peroxynitrite 0.2 mu M stimulated DNA synthesis in PASMC cell by 200 +/- 22% and in PAEC by 137 +/- 4%. DNA synthesis in PAEC and PASMC was also stimulated by the peroxynitrite generator SIN-1 2 mu M. Cell proliferation was accompanied by activation of ERK, which peaked at 15 min and remained elevated for 12 h in PASMC. However peroxynitrite at the concentrations used in this study did not activate the stress pathways p38 mitogen activated protein kinase (MAPK) or Jun N-terminal kinase (JNK). Peroxynitrite-induced proliferation and ERK phosphorylation in PASMC were abolished by the peroxynitrite scavenger ebselen 5 mu M. Peroxynitrite-induced proliferation and extracellular signal-regulated kinase (ERK) phosphorylation in PASMC was prevented by selective inhibitors of MAP kinase kinase (MEK) (U0126 5 mu M, PD98059 50 mu M), Raf-1 (Raf-1 kinase inhibitor 10 mu M), Ras (FPT II and FPT III 10 mu M) and protein kinase C (PKC) (GF109203X 10 mu M). Inhibition of EGF or PDGF receptor signaling using AG-1296. AG-1478 or imatinib prevented peroxynitrite-induced cell proliferation and ERK phosphorylation in PASMC. Peroxynitrite can stimulate proliferation of pulmonary artery cells, involving ERK, PKC and EGF or PDGF receptors.

KW - pulmonary

KW - hypertension

KW - peroxynitrite

KW - endothelial cells

KW - smooth muscle cells

KW - activated protein-kinase

KW - signal-transduction

KW - oxidative stress

KW - nitric-oxide

KW - inhibitor

KW - receptor

KW - pathway

KW - RAF-1

KW - RAS

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U2 - 10.1016/j.pupt.2010.09.003

DO - 10.1016/j.pupt.2010.09.003

M3 - Article

VL - 24

SP - 100

EP - 109

JO - Pulmonary Pharmacology and Therapeutics

JF - Pulmonary Pharmacology and Therapeutics

SN - 1094-5539

IS - 1

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