Regulation Of Atorvastatin On The Ultrastructure And Lipid Metabolism Of AC16 Cardiomyocytes Through Endoplasmic Reticulum Stress Under The Influence Of Alcohol

Objective: To investigate the effect of atorvastatin on the ultrastructure and lipid metabolism of AC16 cardiomyocytes through endoplasmic reticulum stress(ERS)under the influence of alcohol and its mechanism.Methods:(1)The ERS model of AC16 cardiomyocytes under the influence of alcohol was established,and it was verified by detecting the expression of ERS-related factor GRP78 at protein level at different time points(0,12,24 and 48h)using western blot.(2)AC16 cardiomyocytes under the influence of alcohol was treated with different concentrations of atorvastatin(1,10,100 μmol/L),the expression of GRP78 protein was detected to confirm the effect of atorvastatin on ERS.The influence of atorvastatin on the ultrastructure of cardiomyocytes was observed using electron microscope,and through the content determination of lipid metabolism key protein,SREBP-1c,and triglyceride(TG),and the detection of intracellular lipid droplets using oil red O staining,the effect of atorvastatin on the lipid metabolism of cardiomyocytes was explored.Results:(1)After treated with 200 mmol/L ethanol for 12,24 and 48 h,the relative expression of GRP78 in the experimental groups were 0.27 ± 0.03,0.39 ± 0.01 and 0.64± 0.02,respectively,while it was 0.19 ± 0.02 in the control group(0h),indicating that the difference was statistically significant.(2)After treated with different concentrations of atorvastatin(1,10,100 μmol/L)for 48 h,(1)the relative expression of GRP78 in atorvastatin groups(1,10,100 μmol/L)were 0.50 ± 0.04,0.38 ± 0.03 and 0.24 ± 0.01(0.19 ± 0.02 in normal group and 0.64 ± 0.02 in alcohol group),respectively,with statistically significant differences;(2)In alcohol group,AC16 cells were abnormal in morphology,mitochondria decreased significantly in number and increased in size,with the disorganization of mitochondria cristae,and sometimes the disappearance of mitochondria cristae,showing vacuole-like changes.With the increase of atorvastatin concentration,the morphology and mitochondrial structure of AC16 cells tended to be normal.Among them,a large number of mitochondria were observed in 100 μmol/L group,they were oval,with regular mitochondria cristae structure,which were same as normal cell group;(3)The relative expression of SREBP-1c in atorvastatin groups(1,10,100 μmol/L)were 0.47 ± 0.04,0.39 ± 0.03 and 0.31 ± 0.02(0.25 ± 0.02 in normal group and 0.56 ± 0.03 in alcohol group),respectively,with statistically significant differences;(4)The relative expression of TG in atorvastatin groups(1,10,100 μmol/L)were 31.73 ±1.59,26.84 ± 1.63 and 23.11 ± 2.05(21.56 ± 1.78,in normal group and 36.35 ± 2.41 in alcohol group),respectively,and there were statistically significant differences between 10 and 100 μmol/L atorvastatin groups and alcohol group.Conclusion: Atorvastatin can inhibit the expression of ERS-related factor GRP78 under the influence of alcohol,improve the ultrastructure(cell morphology,mitochondria,etc.)and lipid metabolism of cell model with alcoholic cardiomyopathy(ACM).