Cardiomyocytes from postinfarction failing rat hearts have improved ischemia tolerance

Am J Physiol Heart Circ Physiol. 2009 Mar;296(3):H787-95. doi: 10.1152/ajpheart.00796.2008. Epub 2009 Jan 9.

Abstract

Altered myocardial Ca(2+) and Na(+) handling in congestive heart failure (CHF) may be expected to decrease the tolerance to ischemia by augmenting reperfusion Ca(2+) overload. The aim of the present study was to investigate tolerance to hypoxia-reoxygenation by measuring enzyme release, cell death, ATP level, and cell Ca(2+) and Na(+) in cardiomyocytes from failing rat hearts. CHF was induced in Wistar rats by ligation of the left coronary artery during isoflurane anesthesia, after which cardiac failure developed within 6 wk. Isolated cardiomyocytes were cultured for 24 h and subsequently exposed to 4 h of hypoxia and 2 h of reoxygenation. Cell damage was measured as lactate dehydrogenase (LD) release, cell death as propidium iodide uptake, and ATP by firefly luciferase assay. Cell Ca(2+) and Na(+) were determined with radioactive isotopes, and free intracellular Ca(2+) concentration ([Ca(2+)](i)) with fluo-3 AM. CHF cells showed less increase in LD release and cell death after hypoxia-reoxygenation and had less relative reduction in ATP level after hypoxia than sham cells. CHF cells accumulated less Na(+) than sham cells during hypoxia (117 vs. 267 nmol/mg protein). CHF cells maintained much lower [Ca(2+)](i) than sham cells during hypoxia (423 vs. 1,766 arbitrary units at 4 h of hypoxia), and exchangeable Ca(2+) increased much less in CHF than in sham cells (1.4 vs. 6.7 nmol/mg protein) after 120 min of reoxygenation. Ranolazine, an inhibitor of late Na(+) current, significantly attenuated both the increase in exchangeable Ca(2+) and the increase in LD release in sham cells after reoxygenation. This supports the suggestion that differences in Na(+) accumulation during hypoxia cause the observed differences in Ca(2+) accumulation during reoxygenation. Tolerance to hypoxia and reoxygenation was surprisingly higher in CHF than in sham cardiomyocytes, probably explained by lower hypoxia-mediated Na(+) accumulation and subsequent lower Ca(2+) accumulation in CHF after reoxygenation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetanilides / pharmacology
  • Adenosine Triphosphate / metabolism
  • Animals
  • Calcium / metabolism
  • Cell Death
  • Cell Hypoxia
  • Cells, Cultured
  • Disease Models, Animal
  • Heart Failure / etiology
  • Heart Failure / metabolism*
  • Heart Failure / pathology
  • L-Lactate Dehydrogenase / metabolism
  • Male
  • Myocardial Infarction / complications*
  • Myocardial Infarction / metabolism
  • Myocardial Infarction / pathology
  • Myocardial Reperfusion Injury / etiology
  • Myocardial Reperfusion Injury / metabolism
  • Myocardial Reperfusion Injury / prevention & control*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / enzymology
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / pathology
  • Oxygen / metabolism*
  • Piperazines / pharmacology
  • Potassium / metabolism
  • Ranolazine
  • Rats
  • Rats, Wistar
  • Rubidium Radioisotopes
  • Sodium / metabolism
  • Sodium Channel Blockers / pharmacology
  • Sodium-Calcium Exchanger / metabolism
  • Sodium-Potassium-Exchanging ATPase / metabolism
  • Time Factors

Substances

  • Acetanilides
  • Piperazines
  • Rubidium Radioisotopes
  • Sodium Channel Blockers
  • Sodium-Calcium Exchanger
  • Adenosine Triphosphate
  • Sodium
  • Ranolazine
  • L-Lactate Dehydrogenase
  • Sodium-Potassium-Exchanging ATPase
  • Potassium
  • Oxygen
  • Calcium