Corosolic Acid Regulates Nrf2 Inhibiting Ferroptosis To Aattenuate Doxorubicin-Induced H9c2 Cardiomyocytes

ObjectiveTo study the mechanism and effect of Corosolic acid（CRA）on doxorubicin（DOX）-induced H9c2 cardiomyocytes.Methods1.H9c2 cells are random Ly divided into 5 groups : control group,model group（DOX）,positive group（DEX）,treatment group（DOX+CRA）and nrf2 inhibitor group（DOX+CRA+ML385）.2.1 μmo L/L Doxorubicin establishes myocardial injury model in vitro.3.MTT assay detects the effects of Corosolic acid on the toxicity of H9c2 cardiomyocytes.4.MTT assay detects the effects of Corosolic acid on the viability of doxorubicin-induced H9c2 cardiomyocytes.5.Hoechst 33342 staining detects the effects of Corosolic acid on the morphology of doxorubicin-induced H9c2 cardiomyocytes.6.DCFH-DA probe detects the effects of Corosolic acid on the ROS level of doxorubicin-induced H9c2 cardiomyocytes.7.Iron colorimetry detects the effects of Corosolic acid on the Fe²⁺ level of doxorubicin-induced H9c2 cardiomyocytes.8.Annexin V-FITC/PI detects the effects of Corosolic acid on the apoptosis rate of doxorubicin-induced H9c2 cardiomyocytes.9.Western blot detects the effects of Corosolic acid on the protein expression of Bax,Bcl-2,Cleaved caspase3,Nrf2,Gpx4 and Ptgs2 in doxorubicin-induced H9c2 cardiomyocytes.ResultsCompared with the blank group,the H9c2 cardiomyocytes in the model group showed DOX increased the level of Fe²⁺,ROS,and apoptosis,promotoed the protein expression of Bax,Ptgs2 and Cleaved caspase3,and decreased the protein expression of Nrf2,Gpx4 and Bcl-2（P<0.05）.Compared with the model group,the DOX-induced H9c2 cardiomyocytes in the treatment group showed CRA decreased the level of Fe²⁺,ROS,and apoptosis,downregulated the protein expression of Bax,Ptgs2 and Cleaved caspase3,and promotoed the protein expression of Nrf2,Gpx4 and Bcl-2（P<0.05）.Compared with the treament group,the DOX-induced H9c2 cardiomyocytes in the Nrf2 inhibitor group showed ML385 downregulated the protein expression of Gpx4（P<0.05）.Conclusions1.Corosolic acid can alleviate doxorubicin-induced myocardial injury.2.Corosolic acid reduces doxorubicin-induced cardiomyocyte apoptosis by downregulating the protein expression of Bax and Cleaved caspase3,and upregulating the protein expression of Bcl-2.3.Corosolic acid reduces doxorubicin-induced cardiomyocyte ferroptosis by upregulating Nrf2 pathway to promote the protein expression of Gpx4 and inhibit the protein expression of Ptgs2.