Induction of VEGF gene transcription by IL-1 beta is mediated through stress-activated MAP kinases and Sp1 sites in cardiac myocytes

J Mol Cell Cardiol. 2000 Nov;32(11):1955-67. doi: 10.1006/jmcc.2000.1228.

Abstract

Interleukin-1 beta (IL-1 beta) is a multipotent cytokine participating in a variety of cardiovascular diseases. In this study, we examined the effects of IL-1 beta on the expression of vascular endothelial cell growth factor (VEGF) and pursued the molecular mechanisms underlying this effect. Treatment of cultured neonatal rat cardiac myocytes with IL-1 beta increased the levels of VEGF mRNA in a time- and a concentration-dependent manner. These effects were completely abolished by SB203580 and SB202190 (p38 MAPK inhibitors) but not by PD98059 (MEK1 inhibitor), calphostin C (protein kinase C inhibitor), or genistein (tyrosine kinase inhibitor). While IL-1 beta phosphorylated c-Jun N-terminus protein kinase (JNK) rapidly and transiently, the effect of IL-1 beta on p38 mitogen-activated protein kinase (MAPK) was gradual and persistent. Transient transfection assays showed that IL-1 beta increases the transcription from the VEGF promoter. A series of 5;-deletion and site-specific mutation analyses indicated that IL-1 beta as well as overexpression of p38 MAPK and JNK activate VEGF promoter activity through two G+C-rich sequences located at -73 and -62. Electrophoretic mobility shift and supershift assays showed Sp1 and Sp3 proteins specifically bind to the G+C-rich sequences. The half-life of VEGF mRNA was significantly increased in cells treated with IL-1 beta. Together, these results indicate that IL-1 beta induces VEGF gene expression at both transcriptional and post-transcriptional levels, and IL-1 beta evokes p38 MAPK and JNK signalings, which in turn stimulate the transcription of the VEGF gene through Sp1-binding sites. These findings suggest the role of IL-1 beta as a cytokine inducing VEGF in cardiac myocytes, and imply that activation of stress-activated MAP kinases regulate Sp1 sites-dependent transcription.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cells, Cultured / drug effects
  • Cells, Cultured / metabolism
  • Dactinomycin / pharmacology
  • Dinoprost / pharmacology
  • Endothelial Growth Factors / biosynthesis*
  • Endothelial Growth Factors / genetics
  • Endothelin-1 / pharmacology
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Regulation / drug effects*
  • Genistein / pharmacology
  • Heart / drug effects*
  • Imidazoles / pharmacology
  • Interleukin-1 / genetics
  • Interleukin-1 / physiology*
  • JNK Mitogen-Activated Protein Kinases
  • Lipopolysaccharides / pharmacology
  • Lymphokines / biosynthesis*
  • Lymphokines / genetics
  • MAP Kinase Signaling System / drug effects*
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism
  • Myocardium / cytology
  • Myocardium / metabolism*
  • Naphthalenes / pharmacology
  • Nucleic Acid Synthesis Inhibitors / pharmacology
  • Promoter Regions, Genetic
  • Protein Kinase C / antagonists & inhibitors
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Pyridines / pharmacology
  • RNA, Messenger / biosynthesis
  • Rats
  • Rats, Wistar
  • Recombinant Fusion Proteins / physiology
  • Regulatory Sequences, Nucleic Acid
  • Sp1 Transcription Factor / metabolism*
  • Stress, Physiological / enzymology
  • Transcription, Genetic / drug effects*
  • Transfection
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • p38 Mitogen-Activated Protein Kinases

Substances

  • Endothelial Growth Factors
  • Endothelin-1
  • Enzyme Inhibitors
  • Imidazoles
  • Interleukin-1
  • Lipopolysaccharides
  • Lymphokines
  • Naphthalenes
  • Nucleic Acid Synthesis Inhibitors
  • Pyridines
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Sp1 Transcription Factor
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • calphostin complex
  • Dactinomycin
  • Dinoprost
  • Genistein
  • Protein-Tyrosine Kinases
  • Protein Kinase C
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases
  • SB 203580
  • 4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole