The Mechanism of METTL14-mediated m6A Methylation Regulating SIRT6 in Myocardial Ischemia-reperfusion Injury
HE Juan
HUANG Min
XIE Aiping
HE Xiubo
Abstract：Objective:To explore the mechanism of methyltransferase-like protein 14(METTL14)regulating silent information regulator 6(SIRT6)myocardial ischemia-reperfusion(I/R)injury in mice.Methods:Sixty healthy C57BL/6 mice were selected and the myocardial I/R injury model was established by coronary artery ligation.The overexpression plasmid of METTL14 and negative controls[phosphorylated METTL14(p-METTL14),p-NC]were contracted,siRNA of SIRT6 and negative controls(si-SIRT6,si-NC)METTL14 and SIRT6 were overexpressed or inhibited respectively in mice by tail vein injection after being packaged with lentivirus.The mice were randomly divided into the sham operation group(Sham group),the model group(I/R group),the p-METTL14 group,the p-NC group,the p-METTL14+si-SIRT6,and the p-METTL14+si-NC group,with 10 rats in each group.The changes of cardiac function in mice were evaluated by the animal echocardiography system.The myocardial infarction area of mice was determined by Evans blue/2,3,5-triphenyltetrazolium chloride(TTC)staining.The apoptosis level of cardiomyocytes was detected by deoxyribonucleotide terminal transferase-mediated notch end labeling(TUNEL)staining.The expression level of SIRT6 mRNA and its m6A methylation level in mice myocardial tissue were detected by m6A methylation co-immunoprecipitation(MeRIP)-polymerase chain reaction(qPCR)method.The expression of autophagy marker proteins in mitochondria of myocardial tissue was detected by Western Blot.Results:Compared with the Sham group,the left ventricular ejection fraction(EF),short-axis shortening rate(FS),and the expressions of SIRT6 mRNA,obesitation-associated protein(FTO),methyltransferase-like protein 3(METTL3),and ALKB homolog 5(ALKBH5)in myocardial tissue of mice in the I/R group significantly decreased,the myocardial infarction area,the apoptosis rate of myocardial cells,and the relative expression levels and gray values of mitochondrial Parkinson's disease protein(Parkin)and Pent-induced kinase 1(PINK1)proteins in myocardial tissue significantly increased(P＜0.05).Compared with the I/R group,the expression of SIRT6 mRNA in EF,FS and myocardial tissues of mice in the p-METTL14 group and the p-METTL14+si-SIRT6 group significantly increased,the myocardial infarction area,myocardial cell apoptosis rate,FTO,METTL3,ALKBH5,and the relative expression levels and gray values of Parkin and PINK1 proteins in myocardial tissue mitochondria significantly decreased,and the p-METTL14+si-SIRT6 group was superior to the p-METTL14 group(P＜0.05).Conclusion:METTL14 shows protective effect on I/R injury in mice.It may improve the cardiac function of mice,reduce the myocardial infarction area,inhibit myocardial cell apoptosis and promote mitochondrial autophagy in myocardial cells by enhancing the m6A methylation modification of SIRT6,and alleviate myocardial I/R injury in mice.
Keywords：myocardial ischemia-reperfusion injurymethyltransferase-like protein 14METTL14silent information regulator 6SIRT6N6-methyladenosineRNA methylationmitochondrial autophagyexperimental study
Publication Date：2025-06-10
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 1658-1665 )