Ouabain decreases sarco(endo)plasmic reticulum calcium ATPase activity in rat hearts by a process involving protein oxidation Article (Web of Science)

abstract

  • The effect of cardiac glycosides to increase cardiac inotropy by altering Ca2+cycling is well known but still poorly understood. The studies described in this report focus on defining the effects of ouabain signaling on sarcoplasmic reticulum Ca2+-ATPase function. Rat cardiac myocytes treated with 50 μM ouabain demonstrated substantial increases in systolic and diastolic Ca2+concentrations. The recovery time constant for the Ca2+transient, τ[Formula: see text], was significantly prolonged by ouabain. Exposure to 10 μM H2O2, which causes an increase in intracellular reactive oxygen species similar to that of 50 μM ouabain, caused a similar increase in τ[Formula: see text]. Concurrent exposure to 10 mM N-acetylcysteine or an aqueous extract from green tea (50 mg/ml) both prevented the increases in τ[Formula: see text] as well as the changes in systolic or diastolic Ca2+concentrations. We also observed that 50 μM ouabain induced increases in developed pressure in addition to diastolic dysfunction in the isolated perfused rat heart. Coadministration of ouabain with N-acetylcysteine prevented these increases. Analysis of sarcoplasmic reticulum Ca2+-ATPase protein revealed increases in both the oxidation and nitrotyrosine content in the ouabain-treated hearts. Liquid chromatography-mass spectrometric analysis confirmed that the sarcoplasmic reticulum Ca2+-ATPase protein from ouabain-treated hearts had modifications consistent with oxidative and nitrosative stress. These data suggest that ouabain induces oxidative changes of the sarcoplasmic reticulum Ca2+-ATPase structure and function that may, in turn, produce some of the associated changes in Ca2+cycling and physiological function.

authors

  • Kennedy, David J.
  • Vetteth, Sandeep
  • Xie, Miaorong
  • Periyasamy, Sankaridrug M.
  • Xie, Zijian
  • Han, Chi
  • Basrur, Venkatesha
  • Mutgi, Krishna
  • Fedorov, Vladimir
  • Malhotra, Deepak K
  • Shapiro, Joseph I.

publication date

  • 2006

webpage

published in

start page

  • H3003

end page

  • H3011

volume

  • 291

issue

  • 6