Effect Of α-1 Antitrypsin On PI3K/AKT Of Hypoxia/Reoxygenation Of Mouse Cardiomyocytes

Objective:To establish a hypoxia/reoxygenation model of mouse cardiomyocytes,observe the expression of Phosphatidylinositol 3-kinas（PI3K）,Protein Kinase B（AKT）and caspase-3 in mouse cardiomyocytes with hypoxia/reoxygenation,and explore the effects ofα-1 antitrypsin on mouse cardiomyocytes through intervention of PI3K/AKT The effects of hypoxia/reoxygenation.Methods:Subgroup:normal group（Control group）,hypoxia reoxygenation group（H/R group）,α-1 anti trypsin intervention group（A1AT group）,In vitro,mouse cardiomyocytes were cultured with Na₂S₂O₄-induced hypoxia medium after hypoxia and reoxygenation to simulate hypoxia/reoxygenation model,to choose the best intervention concentration of A1AT.CCK-8 was used to detect cell function,optical microscope was used to observe cell morphology changes,expression of PI3K,AKT and downstream molecules were detected by immunofluorescence and Western Blot.SPSS 22.0 was used for data statistics.Results:1.The best time for Na₂S₂O₄to induce hypoxia in cardiomyocytes is 60min,the best concentration for inducing hypoxia is 4mmol/L,the best intervention concentration for A1AT is 20umol/L,and the best intervention time is 4 hours.2.The cardiomyocytes of mice in the Control group are arranged neatly,tightly,fusiform and regular in shape.The arrangement of myocardial cells in the H/R group was irregular,sparser than that in the normal group,and the cells were hypertrophic,partly elliptical or irregular.The arrangement of cardiomyocytes in the A1AT group was more regular and tighter than that in the H/R group,and the hypertrophic cardiomyocytes were significantly reduced.3.Immunofluorescence detection revealed that the expression of PI3K,AKT,and Caspase 3 molecules in the H/R group was Strong positive.Compared with the Control group and the AST group,the difference was statistically significant（P<0.05）.4.WB detection revealed that the immunofluorescence detection found that the expression of PI3K,AKT and caspase-3molecules in the H/R group was up-regulated.Compared with the Control group and the AST group,the difference was statistically significant（P<0.05）.Conclusion:1.PI3K、AKT and caspase-3 expression were up-regulated in cardiomyocyte hypoxia/reoxygenation.2.A1AT can inhibit the expression of PI3K,AKT and caspase-3 in hypoxia/reoxygenation and reduce hypoxia/reoxygenation injury in cardiomyocytes.