Effects of Hypothermia on Astrocytes Death Due to Depolarization or Inhibitors of Glycolysis and Oxidative Phosphorylation / 대한마취과학회지

ABSTRACT

BACKGROUND: There are evidences that cytotoxic cell death occurs first by intracellular sodium entry and then followed by calcium accumulation during ischemic damage. To investigate the protective effect of hypothermia on the sodium induced or energy depletion induced cell death, we studied the relationship of incubation temperature with viability of the cultured astrocytoma cells. METHODS: The survival rate of astrocytoma cells under veratridine and/or iodoacetate(IAA)/carbonylcyanide m-chlorophenylhydrazone (CCCP) treatments was assessed. To measure the cell viability by veratridine or IAA/CCCP, 3-[4,5-dimethylthiazol-2yl]-2,5, diphenyl tetrazolium bromide (MTT) test using ELISA was utilized. Incubation temperature was varied to 27, 30, 37oC. RESULTS: Veratridine (30, 15, 3 M) known to increase intracellular sodium caused cell death. The survival rate was 88.8 1.3, 100.04 3.8, 105 4.5% of control, respectively at 1hr and 80.0 1.72, 90.9 1.68, 97.5 0.9%, of control respectively at 3 hrs after treatment. The survival rate with IAA/CCCP 1.5 mM/20 M or 150 M/2 M was 12.75 0.99, 32.85 2.93, respectively at 1 hr, and 3.1 0.36%, 15.48 1.11, respectively at 3 hrs. Veratridine addition to IAA/CCCP exacerbated cell death as compared with IAA/CCCP alone (6.6 0.43 vs 15.48 1.11). Lowering incubation temperature decreased cell death by veratridine or IAA/CCCP significantly: veratridine treated group revealed 80.0 1.72 % survival rate at 37oC and 94.1 4.0% at 27oC after 3 hrs incubation. IAA/CCCP (150 M/2 M) treated group showed 15.48 1.11% survival rate at 37oC and 39.96 5.20% survival rate at 27oC after 3 hrs incubation. CONCLUSIONS: Cell death caused by veratridine or IAA/CCCP was ameliorated by hypothermic incubation.