Effect of Mangiferin on Myocardial Ischemia-reperfusion Injury in Rats Based on Nrf2-GPX4 Ferroptosis Pathway
Objective:To investigate the protective effect of mangiferin on myocardial ischemia-reperfusion injury(MIRI)in rats.Methods:Sixty healthy male SD rats without specific pathogen(SPF)grade were randomly divided into sham operation group,MIRI group,low-dose mangiferin group,high-dose mangiferin group,and mangiferin +Nrf2 inhibitor(ML385)group,with 12 rats in each group.Except the sham group,the MIRI model was established by ligation of the anterior descending branch of the left coronary artery.After 4 h of reperfusion,serum troponin I(cTnI),lactate dehydrogenase(LDH)and creatine kinase(CK)levels were determined by enzyme-linked immunosorbent assay(ELISA).The cardiac histopathological changes were observed by hematoxylin-eosin(HE)staining.Myocardial cell apoptosis was analyzed by terminal deoxyribonucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)method.Dihydroethidium(DHE)fluorescence assay was used to detect the level of reactive oxygen species(ROS)in myocardial tissue.Superoxide dismutase(SOD)activity,malondialdehyde(MDA),glutathione(GSH),and ferrous ion(Fe2+)contents were detected by colorimetry.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western Blot were used to detect the gene and protein expression of nuclear factor E2-related factor 2(Nrf2),glutathione peroxidase 4(GPX4),ferritin heavy chain 1(FTH1),and long chain fatty acyl-CoA synthetase 4(ACSL4)in myocardial tissue.Results:Compared with sham group,serum levels of cTnI,CK and LDH,cardiomyocyte apoptosis rate,ROS,MDA,Fe2+ content,ACSL4 mRNA and protein levels of rats in MIRI group increased,SOD activity and GSH content,Nrf2,GPX4,FTH1 mRNA and protein levels decreased(P<0.05),cardiomyocytes were swollen,myocardial fibers were disordered,and a large number of inflammatory cells were infiltrated.Compared with MIRI group,serum cTnI,CK and LDH levels,cardiomyocyte apoptosis rate,ROS,MDA and Fe2+ contents,ACSL4 mRNA and protein levels in low-dose mangiferin group,high-dose mangiferin group decreased,SOD activity and GSH content,Nrf2,GPX4,FTH1 mRNA and protein levels increased(P<0.05),cardiomyocyte edema reduced,with a small amount of myocardial fiber rupture and inflammatory cell infiltration.Inhibition of nuclear translocation of Nrf2 by Nrf2 inhibitor ML385 could significantly block the inhibitory effect of mangiferin on cardiac ferroptosis(P<0.05).Conclusion:Mangiferin can protect MIRI by inhibiting ferroptosis through activation of Nrf2-GPX4 axis.
myocardial ischemia-reperfusion injuryferroptosismangiferinnuclear factor-E2-related factor 2glutathione peroxidase 4experimental study
万方数据
维普
