Study on Protective Effect and Mechanism of Astragaloside Ⅳ on D-galactose-induced Primary Cardiomyocytes Apoptosis / 中国药房

ABSTRACT

OBJECTIVE:To study the protective effect and mechanism of astragaloside Ⅳ on D-galactose(D-gal)-induced primary cardiomyocytes apoptosis. METHODS:Wistar neonatal rat primary cardiomyocytes were isolated and cultured. The cardiomyocytes were divided into normal control group(DMEM high glucose medium without serum),D-gal group(DMEM high glucose medium without serum containing 5 g/L D-gal)and astragaloside Ⅳ low-concentration,medium-concentration and high-concentration groups(after cultured with DMEM high glucose medium without serum containing 25,50,100 μg/mL astragaloside Ⅳ for 1 h,and then replaced with DMEM high glucose medium without serum containing corresponding concentration of astragaloside Ⅳ and 5 g/L D-gal). Cardiomyocytes activity was detected by CCK-8 kit after cultured for 48 h.The apoptosis level was detected by Hoechst staining(cultured for 24 h)and flow cytometry(cultured for 48 h). The mRNA expressions of apoptosis related factors(Bcl-2,Bax,Caspase-3)and ANP in cardiomyocytes were detected by RT-PCR after cultured for 24 h. RESULTS:Compared with normal control group,the activity of cardiomyocytes,mRNA level of Bcl-2 and ratio of Bcl-2/Bax were decreased in D-gal group,while apoptosis rate and mRNA levels of Bax,Caspase-3 and ANP were increased, with statistical significance(P<0.05 or P<0.01). Compared with D-gal group,the activity of cardiomyocytes,mRNA level of Bcl-2 and ratio of Bcl-2/Bax were increased in astragaloside Ⅳ groups,while apoptosis rate and mRNA levels of Bax,Caspase-3 and ANP were decreased,with statistical significance(P<0.05 or P<0.01),and in concentration-dependent manner. CONCLUSIONS:Astragaloside Ⅳ can protect D-gal induced primary cardiomyocytes from apoptosis,the mechanism of which may be associated with the increase of Bcl-2/Bax ratio and the decrease of mRNA expressions of Caspase-3 and ANP.