Protective Effect Of Metallothionein 2A On Myocardial Ischemia Reperfusion Injury

Objective Cardiovascular disease is a serious disease that seriously affects human health.Coronary heart disease accounts for the major part of the incidence of cardiovascular disease.Acute myocardial infarction is the leading cause of cardiovascular mortality.Revascularization has been the primary goal of acute myocardial infarction,but reperfusion after ischemia will lead to ischemia-reperfusion injury.Ischemia-reperfusion injury leads to increased myocardial injury,and even irreversible damage.At present,there is still no effective clinical treatment for myocardial ischemia-reperfusion injury.Metallothionein 2A is an important anti-oxidant stress protein,which has anti-oxidative stress and regulates the autophagy and apoptosis.Myocardial Ischemia-reperfusion Injury by using Cobalt Chloride to induce H9c2 cardiomyocytes to Construct Hypoxia / Reoxygenation Model.Construction of stable MT2 A overexpression strain To investigate the role of metallothionein 2A in cardiomyocyte injury induced by hypoxia / reoxygenation and its possible mechanism.Methods H9c2 cells were induced by cobalt chloride to establish hypoxia / reoxygenation model to simulate myocardial ischemia-reperfusion injury.Overexpression of MT2 A protein in H9c2 cardiomyocytes and explore the role of metallothionein 2A in cardiomyocyte injury induced by hypoxia / reoxygenation and its possible mechanism.Grouping: normoxia + control,normoxia + vector,normoxia + MT2 A,hypoxia /reoxygenation + control,hypoxia / reoxygenation + vector,hypoxia / reoxygenation + MT2 A.Fluorescence microscopy and Western blot were used to confirm the MT2 A overexpression stable transfected cells;MTT assay test cell viability;DHE probe loading cells,the expression of ROS detected by flow cytometry;Western blot was used to detect the expression of apoptosis-related proteins Cleaved Caspase-3.Results 1,MT2 A lentiviral vector infection H9c2 cells.Compared with the control group,the fluorescence intensity of vector group and MT2 A group increased significantly.Compared with the control group,the expression level of MT2 A in vector group had no significant change.MT2 A protein expression in MT2 A group was significantly increased than that in vector group(P <0.01).2,Under hypoxia / reoxygenation conditions.Compared with the control group,the cell survival rate of vector group was not changed significantly after hypoxia / reoxygenation treatment.The cell survival rate of MT2 A group increased significantly than that in vector group(P <0.01).3,Under normoxic conditions,compared with the control group,the level of ROS in the vector group was not significantly changed;compared with the vector group,the ROS level in the MT2 A group was down-regulated(P <0.01).Under hypoxia / reoxygenation conditions,compared with the control group,there was no significant change in ROS level in the vector group;compared with the vector group,ROS level in the MT2 A group was significantly decreased(P <0.01).4,Under normoxic conditions,compared with the control group,the level of Cleaved Caspase-3 in the vector group was not significantly changed;compared with the vector group,the level of Cleaved Caspase-3 in the MT2 A group was down-regulated(P <0.01).Under hypoxia / reoxygenation conditions,compared with the control group,the level of Cleaved Caspase-3 in the vector group was not significantly changed;compared with the vector group,the level of Cleaved Caspase-3 in the MT2 A group was down-regulated(P <0.01).Conclusions In the cobalt chloride-induced hypoxia / reoxygenation injury H9c2 cardiomyocyte model,metallothionein 2A increased cardiomyocyte cell viability,decreased ROS levels,decreased the expression of apoptosis-related proteins Cleaved Caspase-3.Metallothionein 2A plays a role in myocardial protection during cardiomyocyte hypoxia / reoxygenation injury,which may be related to its anti-oxidative stress,anti-apoptosis.