Effects Of MiR-29b/c On Proliferation And Angiogenesis Of Ischemic Cardiomyopathy

Ischemic cardiomyopathy is one of the most common heart disease,caused by hypoxia-ischemia.It can lead to myocardial infarction,thus weakening myocardial cell viability and being replaced by inactive cardiomyopathy,which ultimately resulted in myocardial remodeling and heart failure.At present,many biological technologies based on cell and non-cell are used to cure ischemic cardiomyopathy.But none of these are applied to clinic[1-3].MicroRNAs(miRNAs)are endogenous,single-stranded RNA molecules,consisting of approximately 18-25 non-coding nucleotides that exist in various of organisms.The miR-29 family is a conserved family of miRNAs including miR-29a/b/c,playing an important role in kinds of disease[4-7].But less research is reported on the mechanism of miR-29b/c on proliferation and angiogenesis of ischemic cardiomyopathy.The vascular ndothelial growth factor A(VEGFA)is the important regulatory factor in angiogenesis and proliferation.It can activate fibroblast translate into myofibroblast,leading to ventricular remodeling and cardiac dysfunction[8,9].The hypoxia-inducible factor 1?(HIF-1?)is the key transcription factor of oxygen balance.It is proved to play a crucial part in many physiological and pathological processes,such as cell proliferation,metabolism,angiogenesis,migration and apoptosis[10-12].In this study,we chose human cardiac fibroblast cells(HCFs)and human umbilical vein endothelial cells(HUVECs)as objects to probe the effect and mechanism of miR-29b/c on cell proliferation and angiogenesis of ischemic cardiomyopathy,offering a brand-new therapeutic method for ischemic cardiomyopathy.Objective:To probe the effect and mechanism of miR-29b/c on cell proliferation and angiogenesis of ischemic cardiomyopathy and offer a brand-new therapies for ischemic cardiomyopathy.Methods:(1)Cultivate HCFs cells and transfect miR-29b/c mimics and non-specific miRNA to HCFs cells.Then analyze the expression of miR-29b/c by RT-qPCR and verify transfection efficiency.(2)Detect the effect of miR-29b/c on proliferation of HCFs cells by CCK-8 and angiogenesis of HUVECs cells by tube formation.(3)Predict the target genes of miR-29b/c using DIANA-TarBase v7.0 and detect the expression of target genes in mRNA and protein level.(4)Inspect the expression of miR-29b/c on ischemis cardiomyopathy with the help of gene chip.Results:(1)After transfecting miR-29b/c mimics,the expression of miR-29b/c was significantly increased(***P<0.0001).Under the electron microscope,above 90%cells carried fluorescence,suggesting that miR-29b/c transfected successfully into cells.(2)After miR-29b/c mimics transfection,the cells viability was decreased in 48h,72h and 96h(*P<0.05,**P<0.01,***P<0.0001).And the capacity of angiogenesis was reduced by tube formation.(3)Through DIANA-TarBase v7.0 software,downstream target gene,HIF-1? and VEGFA of miR-29b/c were predicted.Besides,form the results of Western Blot technique,miR-29b/c suppressed angiogenesis and cell proliferation through down-regulating the expression of HIF-la and VEGFA.(4)The expression of miR-29b/c were lower in ischemic cardiomyopathy.Conclusion:miR-29b/c suppressed angiogenesis and cell proliferation through down-regulating the expression of HIF-1? and VEGFA.