The effects of trimetazidine on palmiticacid-induced lipotoxic apoptosis in H9C2 cardiomyocytes
Objective:To establish a palmitic acid(PA)-induced apoptosis model of H9C2 cells to verify whether trimetazidine(TMZ)had a protective effect on lipotoxic apoptosis in H9C2 cells and its specific mechanism.Methods:H9C2 cardiomyocytes cultured in vitro were stimulated with PA for different time(0,6,12,24 h)and different concentrations(0,50,100,150,200,300,350 μmol/L)(n=3)when the cell density was 90%to establish a lipotoxic injury model.CCK8 was used to detect cell viability to determine the optimal stimulation conditions,and Western blotting was used to confirm cell apoptosis.There were 6 groups in the experiment,including con trol group(CON),PA(200 μmol/L),CON+TMZ3(10 μmol/L),PA(200 μmol/L)+TMZ1(0.1 μmol/L),PA(200 μmol/L)+TMZ2(1 μmol/L),and PA(200 μmol/L)+TMZ3(10 μmol/L)(n=3).CCK8 was used to detect cell viability and Western blotting was used to determine the expression of apoptosis-related proteins.To observe the changes in intracellular lipid content,the experiment was divided into 4 groups,including control group(CON),PA(200 μmol/L),TMZ3(10 μmol/L),and PA(200 μmol/L)+TMZ3(n=3),which were observed under the microscope by Oil Red O staining.Results:Compared with the CON group,the cell viability in the 200 μmol/L PA group and the 12 h group was significantly decreased(t=15.8,20.82,both P<0.05).CCK8 was used to detect and finally determine that under the conditions of PA 200 μmol/L and 12 h stimulation,H9C2 cells suffered greater lipotoxic damage and the most surviving cells.Compared with the CON group,the expression of Bax protein in the 200 μmol/L PA group was significantly increased(t=3.201,P<0.05),and the expression of Bcl-2 protein in the 150 μmol/L PA group was significantly decreased(t=2.479,P<0.05).Protein detection confirmed cell apoptosis.Compared with the PA stimulation group,the cell activity of the PA+TMZ1 group,PA+TMZ2 group,and PA+TMZ3 group increased(F=420.1,all P<0.05),the expression of apoptosis-related protein Bcl-2 increased(t=6.028,8.952,both P<0.05)and the expression of Bax decreased(t=3.392,4.275,all P<0.05)in the PA+TMZ2 group and PA+TMZ3 group.With the increase of TMZ concentration,the expression of Bax decreased significantly and the expression of Bcl-2 increased significantly(F=3.763,7.548,both P<0.05).Compared with the CON group,the intracellular lipid droplet content of H9C2 cells in the PA group increased significantly.Compared with the PA group,the intracellular lipid droplet content of H9C2 cells in the PA+TMZ3 group de-creased significantly.Conclusion:Palmitic acid stimulation can induce lipotoxic apoptosis in H9C2 cardiomyocytes,and trimetazidine can inhibit palmitic acid-induced lipotoxic apoptosis in H9C2 cells.
万方数据
维普
