Angiotensin II inhibits the Na+-K+ pump via PKC-dependent activation of NADPH oxidase

Am J Physiol Cell Physiol. 2009 Apr;296(4):C693-700. doi: 10.1152/ajpcell.00648.2008. Epub 2009 Feb 4.

Abstract

The sarcolemmal Na(+)-K(+) pump, pivotal in cardiac myocyte function, is inhibited by angiotensin II (ANG II). Since ANG II activates NADPH oxidase, we tested the hypothesis that NADPH oxidase mediates the pump inhibition. Exposure to 100 nmol/l ANG II increased superoxide-sensitive fluorescence of isolated rabbit ventricular myocytes. The increase was abolished by pegylated superoxide dismutase (SOD), by the NADPH oxidase inhibitor apocynin, and by myristolated inhibitory peptide to epsilon-protein kinase C (epsilonPKC), previously implicated in ANG II-induced Na(+)-K(+) pump inhibition. A role for epsilonPKC was also supported by an ANG II-induced increase in coimmunoprecipitation of epsilonPKC with the receptor for the activated kinase and with the cytosolic p47(phox) subunit of NADPH oxidase. ANG II decreased electrogenic Na(+)-K(+) pump current in voltage-clamped myocytes. The decrease was abolished by SOD, by the gp91ds inhibitory peptide that blocks assembly and activation of NADPH oxidase, and by epsilonPKC inhibitory peptide. Since colocalization should facilitate NADPH oxidase-dependent regulation of the Na(+)-K(+) pump, we examined whether there is physical association between the pump subunits and NADPH oxidase. The alpha(1)-subunit coimmunoprecipitated with caveolin 3 and with membrane-associated p22(phox) and cytosolic p47(phox) NADPH oxidase subunits at baseline. ANG II had no effect on alpha(1)/caveolin 3 or alpha(1)/p22(phox) interaction, but it increased alpha(1)/p47(phox) coimmunoprecipitation. We conclude that ANG II inhibits the Na(+)-K(+) pump via PKC-dependent NADPH oxidase activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetophenones / pharmacology
  • Angiotensin II / metabolism*
  • Animals
  • Caveolin 3 / metabolism
  • Enzyme Activation
  • Enzyme Inhibitors / pharmacology
  • Free Radical Scavengers / pharmacology
  • Male
  • Membrane Potentials
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / enzymology*
  • NADPH Oxidases / antagonists & inhibitors*
  • NADPH Oxidases / metabolism
  • Peptides / pharmacology
  • Polyethylene Glycols / pharmacology
  • Protein Kinase C-epsilon / antagonists & inhibitors
  • Protein Kinase C-epsilon / metabolism*
  • Rabbits
  • Signal Transduction* / drug effects
  • Sodium-Potassium-Exchanging ATPase / antagonists & inhibitors*
  • Sodium-Potassium-Exchanging ATPase / metabolism
  • Superoxide Dismutase / pharmacology
  • Time Factors

Substances

  • Acetophenones
  • Caveolin 3
  • Enzyme Inhibitors
  • Free Radical Scavengers
  • Peptides
  • Angiotensin II
  • Polyethylene Glycols
  • acetovanillone
  • Superoxide Dismutase
  • polyethylene glycol-superoxide dismutase
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • Protein Kinase C-epsilon
  • Sodium-Potassium-Exchanging ATPase