Effect of apoptosis inducing factor on promoting apoptosis of hypertrophic cardiomyocytes induced by hypoxia/reoxygenation / 解放军医学杂志

ABSTRACT

Objective To evaluate the effect and significance of apoptosis inducing factor (AIF) on promoting apoptosis of hypertrophic cardiomyocytes induced by hypoxia/reoxygenation. Methods Cardiomyocytes of neonatal Kunming strain rats were isolated and cultured. Cardiomyocyte hypertrophy was induced by angiotensinⅡ (0.1?mol/L for 12h), and the cells were cultured under the condition of hypoxia and reoxygenation. The cultured cardiomyocytes were then divided into seven groups hypertrophic cardiomyocyte control (HC) group, hypoxia for 8h (H8h) group, hypoxia for 12h (H12h) group, hypoxia for 8h and reperfusion for 4h (H8h/R) group, hypoxia for 12h and reperfusion for 4h (H12h/R) group, hypoxia for 12h+siRNA transfection (H12h+siRNA) group, and hypoxia for 12h and reperfusion for 4h+siRNA transfection (H12h/R+siRNA) group. Cardiomyocytes were transfected with AIF siRNA. Apoptosis was detected with Hoechst 33258 staining. The AIF mRNA and protein expressions were detected by RT-PCR and Western blotting. Results The levels of AIF mRNA and protein expression were significantly higher in H8h group and H12h group (mRNA 0.52?0.04, 0.85?0.10, respectively; protein 2.07?0.15, 3.12?0.19, respectively) compared with that in HC group (mRNA 0.29?0.08; protein 1.00?0.04, P0.05). The percentage of apoptosis in H12h/R+siRNA group (24.90%?3.90%) was significant higher than that in H12h/R group (14.50%?1.32%, P