Postconditioning prevents apoptotic necrotic and autophagic cardiomyocyte cell death in culture.

ABSTRACT

In the paper the data concerning the possibility of the reproduction of the postconditioning phenomena in the cardyomicyte culture are presented. Primary cultures of cardiomyocytes from neonatal rats underwent 30 minutes of anoxia followed by 60 minutes of reoxygenation. Three different models of postconditioning were used 3 cycles of 1, 3, or 5 minutes of reoxygenation followed by 1, 3, or 5 minutes of anoxia, respectively. The percentage of living, necrotic, and apoptotic cells were determined by staining with Hoechst 33342 and propidium iodide. Autophagy was demonstrated by the staining of vacuolar structures in vivo by monodansyl cadaverine. After anoxia and reoxygenation the amount of living, necrotic and apoptotic cells were 79 +/- 1.5, 7.8 +/- 0.9 and 13 +/- 1.5 %, respectively (in unstimulated cell culture 90 +/- 0.8, 3.3 +/- 0.3, and 5.5 +/- 0.7, P < 0.0001 for all). Postconditioning with 1 min anoxia 3-fold increased the amount of living cells and decreased the number of necrotic and apoptotic cells (P = 0.002, P = 0.02 and P = 0.043 respectively). Postconditioning with cycles of 3 and 5 minutes had a gradually reduced effect compared to cycles of 1 minute. The percentage of autophagic cells in control cell culture was 4.3 +/- 0.3%. This number increased after anoxia-reoxygenation to 14 +/- 0.8%, and was reduced by postconditioning (P < 0.001). The data obtained indicate that postconditioning is one of the effective methods of cardioprotection and could effectively decrease the amount of cardiomyocytes with traits of programmed or non-programmed cell death.