MiR-153-3p Regulates Mitochondrial Division And Cardiac Hypertrophy By Targeting Mfn1

Objective The pathological process of cardiac hypertrophy and the dynamic imbalance of mitochondrial reticular structure-mitochondrial division and fusion are closely related,but even if the mitochondrial division and fusion disorder affect the occurrence and development of various diseases,the mechanism of its involvement in the pathological process of cardiac hypertrophy is still unclear.MicroRNAs(miRNAs)are a class of short-chain non-coding RNAs that promote the expression and translation of a variety ofgenes,thereby regulating various disease processes.MiR-153 has previously been shown to have anti-tumor effects in cancer,especially miR-153-3p has been reported in cancer,including medullary thyroid carcinoma,papillary thyroid cancer,esophageal squamous cell carcinoma,and breast cancer.Although several targets have been identified in cancer cells,the role of miR-153-3p in cardiomyocytes remains to be fully elucidated.Mitochondrial division and fusion are mediated by conserved proteins on the mitochondrial membrane,which hydrolyzegTP to promote the exchange of mitochondrial DNA(mtDNA)proteins and metabolites.Mitochondrial fusion regulates mitochondrial morphogenesis and regulates the progression of multiple diseases.The dynamic balance of mitochondrial network structure is regulated by mitofusin 1/2(Mfn1 / 2),optic nerve atrophin(OPA1)and kinetics Regulation of the protein(Drp1)plays an important role in mitochondrial structure and function.For example,it has been reported that unbalanced processing of OPA1 results in heart failure in mice.In addition,Drp1 can promote the pathogenesis of hypertensive cardiac hypertrophy through ROS production.Therefore,mitochondria are particularly important in cardiomyocytes.In addition,Mfn1 has been shown to be a key player in mediating mitochondrial fusion and morphology in mammalian cells.What is necessary to study the relationship between Mfn1 and cardiac hypertrophy.Therefore,this study will explore the molecular mechanisms of miR-153-3p and MFN1 in cardiomyocyte hypertrophy and mitochondrial division.Methods Our study was performed on primary cardiomyocytes isolated from C57 suckling mice.The cardiomyocyte hypertrophy model was treated by isoproterenol(ISO)treatment,transfected with MFN1 siRNA,miR-153-3p antagomir observed for MFN1 or miR Effect of-153-3p on myocardial hypertrophy and mitochondrial division in cardiomyocytes.The surface area of myocardial cells was detected by phalloidin fluorescence staining.The mitochondrial division of cardiomyocytes was observed by Mito-tracker staining technique.The expression level of MFN1 was detected by western blot.The expression of miRNA and hypertrophic markers was detected by qRT-PCR.Double luciferase reporter The assay was used to detect the targeting effect of miR-153-3p on MFN13’UTR.Result 1.After ISO treatment for 18 h and 24 h,the cell surface area increased,and the expression of atrial natriuretic factor(ANF)and myosin heavy chain(β-MHG)gene was increased.Mitochondrial division occurs in induced cardiomyocytes.2.After ISO treatment for 0h,8h,18 h and 24 h,the expression level of miR-153-3pgradually decreased in a time-dependent manner.RNAhybrid software predicts that MFN1 3’UTR is a potential binding site for miR-153-3p,and decreased expression of miR-153-3p can increase ISO-induced decreased MFN1 levels,and conversely,increase endogenous miR-153-3p Expression can reduce the expression level of MFN1.The dual luciferase reporter assay demonstrated that miR-153-3p indeed inhibits the translational level of the 3’ UTR of MFN1.3.Knockdown of miR-153-3p levels can inhibit cardiomyocyte hypertrophy and mitochondrial division.4.Increasing the expression of MFN1 inhibits ISO-induced cardiomyocyte hypertrophy and mitochondrial division.5.In cardiomyocytes,knockdown of miR-153-3p expression reverses ISO-induced decreased MFN1 levels and decreases mitochondrial division and hypertrophy,whereas MFN1 siRNA attenuates these effects by miR-153-3p antagomir.Conclusion Cardiac hypertrophy and mitochondrial division can occur in myocardial cells under ISO-induced hypertrophy,and this process is associated with up-regulated expression of miR-153-3p.Down-regulation of MFN1 expression is associated with up-regulation of miR-153-3p expression.Regulation of miR-153-3p levels inhibits cardiac hypertrophy and mitochondrial division.miR-153-3p is involved in the regulation of cardiac hypertrophy and mitochondrial division by targeting MFN1.This study demonstrates that the miR-153-3p-MFN1 signaling axis regulates the development of cardiac hypertrophy by participating in the regulation of mitochondrial reticular formation.The specific conclusions are as follows:1.ISO induced myocardial hypertrophy accompanied by mitochondrial division of cardiomyocytes.2.In mouse primary cardiomyocytes,knockdown of miR-153-3p inhibits ISO-induced cardiomyocyte hypertrophy and mitochondrial division.3.Overexpression of MFN1 in mouse primary cardiomyocytes inhibits ISO-induced cardiomyocyte hypertrophy and mitochondrial division.4.At the animal level,knockdown of miR-153-3p inhibits ISO-induced cardiomyocyte hypertrophy and mitochondrial division.5.In mouse primary cardiomyocytes,miR-153-3p regulates ISO-induced cardiomyocyte hypertrophy and mitochondrial division by modulating MFN1 levels.