Study On The Mechanism Of Targeting MiR-34a/SIRT1 Axis To Regulate DCM By Lnc RNA OIP5-AS1

ObjectiveThe expression of long non-coding RNA OIP5-AS1(Lnc RNA OIP5-AS1)and micoRNA-34a(miR-34a)in Diabetic cardiomyopathy(DCM)and their correlation were fully explored.To explore the correlation between miR-34a and SIRT1 in the pathological development of DCM and the regulation of cell activity and oxidative stress.Materials and Methods1.15 DCM patients treated in our hospital were selected as the experimental group(January 2017 to December 2018),and 15 non DCM patients were randomly selected as the control group;In the morning,4.5ml peripheral venous blood was taken from each subject in the two groups.The expression levels of LNC maoip5-asl and miR-34a mRNA in the serum of the two groups were detected by RT qPCR,and their correlation was analyzed.2.Rat cardiomyocytes H9c2 were selected as the research object.H9c2 cells were incubated with normal(5 mM)high glucose solution(10,20,30 mM)for 24 hours and then collected.Rat cardiomyocytes were incubated with high glucose solution(30 mM)and collected.The mechanism of Lnc RNA OIP5-AS1 targeting miR-34a regulating DCM cell proliferation and oxidative stress response was determined.3.The relevant transfected agents miR-NC,miR-34a mimics,inhibitor NC and miR-34a inhibitor were transfected into rat cardiomyocytes respectively to determine the mechanism of miR-34a/SIRT1 axis in DCM.4.Data analysis of cell proliferation and oxidative stress was conducted to determine the mechanism of the regulation of DCM by the Lnc RNA OIP5-AS1 targeting microRNA-34a/SIRT1 axis.Results1.The expression of Lnc RNA OIP5-AS1 in DCM patients was lower than that in the control group.On the contrary,the expression of miR-34a was higher than that in the control group.Poisson distribution showed that there was a negative correlation between them,and the correlation coefficient R2=0.861,the difference was statistically significant(P<0.0001).2.With the increase of glucose concentration,the expression of Lnc RNA OIP5-AS1 decreased,showing a dose-dependent characteristic,and the difference was statistically significant(P<0.05).The Lnc RNA OIP5-AS1 was knocked out in myocardial endothelial cells.When the Lnc RNA OIP5-AS1 was overexpressed,the activity of cells was increased,the expression levels of ROS and MDA were decreased,and SOD was increased,with statistical significance(P<0.05).Overexpression of Lnc RNA OIP5-AS1 in cells can decrease the expression of miR-34a.TargetScan software for data analysis found that miR-34 a mRNA 3 ’noncoding region predicted a OIP5-AS1 binding sites and applications about transfection technology will different combinations of wild type and mutant pGL3OIP5-AS1-UTR expression vector transfection into myocardial cell.Enhancing the expression level of this domain by constructing mutant cell lines can effectively reduce the expression level of miR-34a and reduce the DCM phenotype to a certain extent.3.The overexpression of miR-34a in H9c2 cardiomyocytes significantly decreased the activity of cells,increased the levels of ROS and MDA,and decreased the level of SOD(P<0.05).When miR-34a was overexpressed in H9c2 cardiomyocytes,the expression of SIRT1 protein decreased,and the difference was statistically significant(P<0.05).Overexpression of SIRT1 could increase the activity of cardiomyocytes,reduce the levels of ROS and MDA and increase the level of SOD.A SIRT1 binding site was predicted in the 3 ’noncoding region of miR-34a mRNA.The mutant cell line of SIRT1 binding site was constructed and detected.It was found that the mutation of this site could significantly improve the activity of cardiomyocytes,indicating that SIRT1 site may be the key site for the interaction between miR-34a and Lnc RNA OIP5-AS1.4.Lnc RNA OIP5-AS1 overexpression in H9c2 cardiomyocytes increased SIRT1 expression,and miR-34a mimic could inhibit this trend.The overexpression of Lnc RNA OIP5-AS1 in cardiomyocytes significantly promoted cell activity,decreased the concentrations of ROS and MDA,and increased the expression of SOD.The mimic of miR-34a could inhibited this trend,with statistical significance(P<0.05).Conclusions1.In the pathological development of DCM,the low expression of Lnc RNA OIP5-AS1 and the high expression of miR-34a both promoted the disease development of DCM.The negative correlation between the two expressions was obvious.2.In the development of DCM pathology,Lnc RNA OIP5-AS1 targeted miR-34 a/SIRT1 shaft by influencing the mitochondrial activity of myocardial cells,and oxidative stress to adjust and control the condition:when over expression the Lnc RNA OIP5-AS1,reduced miR-34a expression but increased SIRT1 expression,accordingly cell activity increased,and reduced mitochondrial function disorder,oxidative stress levels slow,inhibited the development of the disease,clinical must be inspired.