A Preliminary Research On LncRNA Involved In The Regulation Of Cell Necrosis To Participate In Myocardial Ischemia-reperfusion Injury Process

Objective:The purpose of this article is to study the roles of lncRNA in H₂O₂-induced cardiomyocyte necrosis.To explore the mechanism of lncRNA regulating myocardial ischemia-reperfusion injury may provide a new target for the treatment of ischemic heart disease.Methods:?1?Construction of mouse cardiomyocyte necrosis model induced by H₂O₂.?2?The target lncRNA was screened by gene chip and RT-qPCR method.?3?The overexpression vector or inhibitory vector of the lncRNA was constructed,and after successful sequencing,the plasmid was extracted for use.?4?Plasmids were transfected and tested for the effect of lncRNA on cell necrosis.?5?To examine the expression of mfn1 in myocardial cell necrosis.The expression of mfn1 mRNA was detected by RT-qPCR,and the expression of mfn1 protein was detected by Western blot.?6?Exploring the relationship between the target lncRNA and mitochondrial fusion protein mfn1.Results:?1?According to the results of PI detection,600umol/L was selected as the concentration of H₂O₂-induced necrosis model of cardiomyocytes.?2?Determine lncRNA NONMMUT026915 as the objective lncRNA of this experiment.?3?After transfecting the plasmids,the result of RT-qPCR showed that the expression of lncRNA NONMMUT026915 in group shRNA was down?P<0.05?.?4?After transfection,the cells were treated with H₂O₂,and the results of PI detection showed that the cell necrosis was reduced in shRNA+H₂O₂ group compared with that of pLKO.1 puro+H₂O₂ group?P<0.05?.?5?H9C2 cells were treated with H₂O₂,and the mRNA and protein expression of mfn1 in the treated group were all down-regulated?P<0.05?.?6?After transfection,the expression of mRNA and protein of mfn1 in shRNA group was up-regulated?P<0.05?.The transfected cells were treated with H₂O₂,and compared with the pLKO.1 puro+H₂O₂group,the expression of mfn1 protein in the shRNA+H₂O₂ group was up-regulated?P<0.05?.Conclusion:The results showed that the expression of lncRNA NONMMUT026915 was up-regulated in mouse model of ischemia-reperfusion injury and the model of cardiomyocyte necrosis induced by H₂O₂.H9C2 cells were used to simulate cardiac myocytes,and the inhibition of the expression of lncRNA NONMMUT026915 could reduce the necrosis of cells induced by H₂O₂ and protect the cardiac myocytes.Mfn1 has the function of promoting mitochondrial fusion and inhibiting cell necrosis,and the expression of mfn1 in H₂O₂-induced cardiomyocyte necrosis model was down-regulation.In the cardiomyocyte necrosis model,inhibition the expression of lncRNA NONMMUT026915 will increase the expression level of mfn1.Therefore,lncRNA NONMMUT026915 plays an important role in cardiomyocyte necrosis,and its target protein may be mfn1.The role of lncRNA NONMMUT026915 in myocardial necrosis may provide a new target for the clinical treatment of ischemic heart disease.