Angiotensin II induces nuclear factor- kappa B activation in cultured neonatal rat cardiomyocytes through protein kinase C signaling pathway

J Mol Cell Cardiol. 2000 Oct;32(10):1767-78. doi: 10.1006/jmcc.2000.1211.


Rat neonatal ventricular cardiomyocytes (RNVM) possess G protein-coupled AT(1)receptors for angiotensin II (AngII) that activate multiple intracellular pathways. To elucidate potential signaling mechanisms involved, we focussed on the nuclear transcription factor-kappa B (NF- kappa B) in RNVM culture. Using specific antibody to NF- kappa Bp65, immunolocalization of NF- kappa B was cytoplasmic in unstimulated cardiomyocytes, whereas NF- kappa B was translocated into the RNVM nucleus in response to AngII. This translocation was inhibited in the presence of calphostin C, a specific inhibitor of protein kinase C (PKC). Western blot analysis showed an increase of NF- kappa B in AngII-stimulated cardiomyocyte nuclear extracts as compared to controls. Biomolecular interaction analysis (BIA analysis) of NF- kappa B activation showed that only AngII-nuclear extracts bound to NF- kappa B consensus sequence with a high degree of affinity. This DNA-binding capacity was completely lost in calphostin C-treated cells. At transcriptional level in RNVM, AngII mediates the upregulation of matrix gelatinase (MMP-9), which is totally inhibited by calphostin C treatment. In conclusion, cardiomyocyte nuclear NF- kappa B translocation in response to Ang II via PKC pathway activates cardiomyocyte-specific transcription of MMP-9 and may activate transcription from responsive genes which are involved in cardiac hypertrophy process and/or cardiac remodeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin II / pharmacology*
  • Animals
  • Animals, Newborn
  • Blotting, Western
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • DNA / metabolism
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Immunohistochemistry
  • Matrix Metalloproteinase 9 / metabolism
  • Microscopy, Confocal
  • Muscles / cytology*
  • Muscles / enzymology
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Naphthalenes / pharmacology
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism*
  • Rats
  • Rats, Wistar
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Surface Plasmon Resonance
  • Time Factors
  • Transcription Factor RelA
  • Transcription, Genetic
  • Up-Regulation


  • Enzyme Inhibitors
  • NF-kappa B
  • Naphthalenes
  • Transcription Factor RelA
  • Angiotensin II
  • DNA
  • Protein Kinase C
  • Matrix Metalloproteinase 9
  • calphostin C