Resisitin Induces H9C2 Cardiomyocyte Hypertrophy Through Ferroptosis

Objective:Through intervention of resistin induced hypertrophy model of H9C2cardiomyocytes with Ferrostatin,the surface area of cardiomyocytes,total protein synthesis of single cells,the expression of cardiomyocyte hypertrophy markers BNP andβ-MHC were detected by real-time fluorescence quantitative PCR,and the expression of iron death related proteins GPX4,ACSL4 and COX2 were detected by Western blot.To investigate whether resistance hormone induces hypertrophy of H9c2cardiomyocytes through iron necrosis,and to explore a new method for effective prevention and treatment of hypertrophy of H9c2 cardiomyocytes,so as to provide theoretical basis for the protection of organ function during perioperative period.Methods:In this study,H9c2 rat cardiomyocytes,which are subclonal cell lines derived from embryonic rat heart tissue,were cultured with DMEM containing 10%FBS,and the culture medium was changed every 2-3 days.When the cells reached 70%to 80%,they were subcultured.Under the microscope,H9c2 cardiomyocytes appear round or oval.Well growing cells were taken for the experiment.1x10~5cardiomyocytes were inoculated into 6-well plates,and 3m L DMEM containing 10%FBS was added for culture.After cell adhesion,3ml DMEM containing 0.1%FBS was changed to starve for 16h.The experiment was divided into four groups,which were divided into Con group according to different experimental groups:control group,after starving cells,0.1%FBS DMEM was changed into 3m L culture for 48h;Res group:resistin group,after starving cells,the resistin concentration was 50ng/m L DMEM containing0.1%FBS 3m L for 48h;Res+FER-1 group:after starvation,cells were changed to1μM fer-1 containing 0.1%FBS for 3m L culture for 16h,and then changed to 50ng/m L resistin containing 0.1%FBS for 3m L culture for 32h.Fer-1 group:After starvation,the cells were cultured with 0.1%FBS 3m L DMEM of 1μM FER-1 for 16h,and then with 0.1%FBS 3m L DMEM for 32h.After 48h culture,H9C2 cells in 4groups were photographed,and the cell surface area was analyzed by Image J software.Cell count was collected,total protein content was measured by BCA method,total protein synthesis of single cell was calculated,and the expression levels of myocardial hypertrophy markers BNP andβ-MHC were detected by real-time fluorescence quantitative PCR.The expression levels of iron death related proteins GPX4,ACSL4 and COX2 were detected by Western blot.Results:Compared with Con group,the myocardial cell surface area,single-cell protein synthesis,the relative expression levels of myocardial cell hypertrophy markers,and the relative expression levels of iron death-related proteins ACSL4 and COX2 were significantly increased in Res group,while the expression levels of GPX4 were significantly decreased,with statistical significance(P<0.05).Compared with Res group,myocardial cell surface area,single-cell protein synthesis,myocardial hypertrophy markers and relative expression levels of iron death related proteins in Res+FER-1 group were significantly decreased,with statistical significance(P<0.05).Conclusion:Resistin can induce hypertrophy of H9C2 cardiomyocytes.Ferroptosis occurred during resistin-mediated hypertrophy of H9c2 cardiomyocytes.The hypertrophy degree of H9c2 myocardial cells could be alleviated by Ferrostatin to a certain extent.Resistin induces hypertrophy of H9C2 cardiomyocytes through the pathway of Ferroptosis.