Selective Inhibition of Inward Rectifier K + Channels (Kir2.1 or Kir2.2) Abolishes Protection by Ischemic Preconditioning in Rabbit Ventricular Cardiomyocytes Journal Articles uri icon

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abstract

  • Volume regulatory Cl − channels are key regulators of ischemic preconditioning (IPC). Because Cl − efflux must be balanced by an efflux of cations to maintain cell membrane electroneutrality during volume regulation, we hypothesize that I K1 channels may play a role in IPC. We subjected cultured cardiomyocytes to 60-minute simulated ischemia (SI) followed by 60-minute of simulated reperfusion (SR) and assessed percent cell death using trypan blue staining. Ischemic preconditioning (10-minute SI/10-minute SR) significantly ( P <0.0001) reduced the percent cell death in nontransfected cardiomyocytes [IPC CM 18.0±2.1% versus control (C CM ) 48.3±1.0%]. IPC protection was not altered by overexpression of the reporter gene (enhanced green fluorescent protein, EGFP). However, overexpression of dominant-negative Kir2.1 or Kir2.2 genes using adenoviruses (AdEGFPKir2.1DN or AdEGFPKir2.2DN) encoding the reporter gene EGFP prevented IPC protection [both IPC CM +AdEGFPKir2.1DN 45.8±2.3% (mean±SEM) and IPC CM +AdEGFPKir2.2DN 47.9±1.4% versus IPC CM ; P <0.0001] in cultured cardiomyocytes (n=8 hearts). Transfection of cardiomyocytes with AdEGFPKir2.1DN or AdEGFPKir2.2DN did not affect cell death in control (nonpreconditioned) cardiomyocytes (both C CM + AdEGFPKir2.1DN 45.8±0.7% and C CM +AdEGFPKir2.2DN 46.2±1.3% versus C CM ; not statistically significant). Similar effects were observed in both cultured (n=5 hearts) and freshly isolated (n=4 hearts) ventricular cardiomyocytes after I K1 blockade with 20 μmol/L BaCl 2 plus 1 μmol/L nifedipine (to prevent Ba 2+ uptake). Nifedipine alone neither protected against ischemic injury nor blocked IPC protection. Our findings establish that I K1 channels play an important role in IPC protection.

authors

  • Diaz, Roberto J
  • Zobel, Carsten
  • Cheol Cho, Hee
  • Batthish, Michelle
  • Hinek, Alina
  • Backx, Peter H
  • Wilson, Gregory J

publication date

  • August 6, 2004

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