Protective effects of NADH on cardiac fibroblasts apoptosis induced by hydrazine / 解放军医学杂志

ABSTRACT

Objective To explore the protection effects of the reduced form of nicotinamide-adenine denucleotid(NADH) on cardiac fibroblasts apoptosis induced by hydrazine in vitro.Methods Cardiac fibroblasts isolated from neonate rats were divided into four groups.In NADH pretreatment before hydrazine treatment group,the cardiac fibroblasts were cultured first in the medium containing 400?g/ml NADH for 2 hours,and then cultured in the medium containing 400?g/ml NADH plus 2mmol/L hydrazine for 72 hours;in hydrazine treatment group,the cardiac fibroblasts were cultured first in normal medium containing 400?g/ml NADH for 2 hours and then cultured in the medium containing 2mmol/L hydrazine for 72 hours;in control group,the cardiac fibroblasts were cultured only in normal medium for 72 hours;and in NADH pretreatment control group,the cardiac fibroblasts were cultured in medium containing 400?g/ml NADH for 72 hours.After the treatments mentioned above,the changes in nuclear of apoptotic cardiac fibroblasts as stained by Hoechst33258 were observed under fluorescence microscope,the apoptotic and necrosis rates of cardiac fibroblasts stained by AnnexinⅤ/PI were assayed by flow cytometry,the mitochondria membrane potential of cells stained by rhodamine123 was also assayed by flow cytometry.Results Hydrazine could induce apoptosis of the cardiac fibroblasts.After NADH pretreatment,the number of apoptotic cardiac fibroblasts with pyknosis of nuclear was lowered,the rates of apoptosis and necrosis decreased,and the mitochondria membrane potential of cells was elevated.Conclusion NADH pretreatment could reduce the apoptosis induced by hydrazine by improving the function of mitochondria.