Effect Of Atorvastatin Calcium Combined With Amlodipine Besylate On Apoptosis Of Cardiomyocytes Induced By Fluctuating High Glucose

ObjectiveCardiomyocytes apoptosis is a main factor leading to heart failure.It was reported that atorvastatin calcium(ATVC)plays an inhibiting role in apoptosis,and amlodipine besylate(AB)could improve the function of cardiac myocytes.However,whether they play a direct or indirect role remain elusive,and underlying molecular mechanism need to further studyb.In this study,H9C2 rat cardiomyocytes were cultured with fluctuating high glucose(FHG)to establish cardiomyocyte apoptosis model,and the effects of atorvastatin calcium and amlodipine besylate on the apoptosis of cardiomyocytes induced by fluctuating high glucose were observed,and the differential expression of apoptosis inhibitor BCL2 was detected.This study provides a new experimental basis for understanding the role of atorvastatin calcium combined with amlodipine besylate in anti apoptosis and for clinical treatment of coronary heart disease.MethodsH9C2 rat cardiomyocytes were cultured in DMEM medium with 10% fetal bovine serum and 1% penicillin.The cells were divided into 5 dishes after passage.The blank control group,the fluctuating high glucose stimulation group,the ATVC culture combined with fluctuating high glucose stimulation group,the AB combined with fluctuating high glucose stimulation group,and the combination of ATVC and AB combined with fluctuating high glucose stimulation group MTT analysis was used to detect the relative survival rate of the cells,Annexin V-FITC / PI double staining cell apoptosis was used to detect the apoptosis,the total proteins of each group were extracted,and the expression level of apoptosis inhibitory protein BCL2 was detected by Western blot.Results1.MTT method was used to detect the relative viability of the cells.It was found that the number of cells decreased significantly under the stimulation of fluctuating high glucose(P < 0.0001).When ATVC or AB was added alone,the effect of fluctuating high glucose on cell number was alleviated(P < 0.0001).Combination of ATVC and AB alleviate the decrease effect by FHG to a greater extent There was significant difference between the two groups(P < 0.0001).2.Annexin V-FITC / PI analysis showed that the apoptosis of cardiomyocytes increased significantly in the early,middle and late period and the whole period under the stimulation of fluctuating high glucose(P < 0.0001).The apoptosis of cardiomyocytes induced by fluctuating high glucose(P < 0.0001)was significantly reduced by adding ATVC or AB alone(P < 0.0001)Combination of ATVC and AB was more resistant to apoptosis induced by FHG stimulation(P < 0.01)3.FHG significantly reduced the expression of BCL2(P < 0.01).Although ATVC alone or AB could partially restore the expression of BCL2,they were not statistically significant(P = 0.024,P = 0.099,P > 0.05).Combination of ATVC and AB significantly restored the BCL2 decreased by fluctuation high glucose(P < 0.05)Conclusions 1.ATVC combined with AB has synergistic protective effect on myocardial cell injury induced by FHG.2.ATVC combined with AB has synergistic inhibition on cardiomyocytes apoptosis induced by fluctuating high glucose.3.The protective mechanism of ATVC combined with AB on FHG induced cardiomyocyte injury may be related to the synergistic inhibition of regulating BCL2 level.