Effect of LncRNA H19 Targeting miR-766-3p on Hypoxia Reoxygenation Induced Apoptosis of Rat Myocardial Cell Line H9c2
XING Zhi
LI Hui
QIU Haitang
DILIHUMAER·Abulaiti
ZHU Mengbai
GAO Ying
Abstract：Objective:To observe the effect of long-chain non-coding ribonucleic acid(lncRNA)H19 on hypoxia-reoxygenation-induced apoptosis of the rat myocardial cell line H9c2,and to explore its targeted regulatory effect on micro ribonucleic acid(miR-766-3p).Methods:The rat myocardial cell line H9c2 was cultured and grouped.Except for the blank group and normal group,the LncRNA H19 upregulation group,LncRNA H19 upregulation control group,LncRNA H19 downregulation group,LncRNA H19 downregulation control group,miR-766-3p upregulation group,miR-766-3p upregulation control group,miR-766-3p downregulation group,and miR-766-3p downregulation control group were transfected with pcDNA3.1-H19,pcDNA3.1-control,si-H19,si-NC,miR-766-3p mimics,miR mimics NC,miR-766-3p inhibitor,and miR inhibitor NC,respectively.Except for the normal group,hypoxia lasted for 24 hours,and reoxygenation lasted for 6 hours.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to determine the expression levels of LncRNA H19 and miR-766-3p in each group.Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)was used to detect the apoptosis rate of cells in each group.RT-qPCR was used to detect the expression levels of exchange protein directly activated by cAMP 1(Epac-1),B-lymphomatoma-2(Bcl-2),and Caspase-3 in each group,and Western Blot was used to detect the expression levels of the corresponding proteins in each group.The dual luciferase reporter gene assay was used to investigate the relationship between the lncRNA H19 and miR-766-3p.Results:Compared with the normal group,the expressions of LncRNA H19 and Epac-1 in the blank group decreased(P＜0.05),while the expressions of miR-766-3p,Bcl-2,Capase3,and cell apoptosis rate increased(P＜0.05).Compared with the blank group,the LncRNA H19 upregulation group showed an increase in the expressions of LncRNA H19 and Epac-1(P＜0.05),and a decrease in the expressions of miR-766-3p,Bcl-2,Caspase 3,and cell apoptosis rates(P＜0.05).In the LncRNA H19 downregulation group,the expressions of LncRNA H19 and Epac-1 decreased(P＜0.05),while the expressions of miR-766-3p,Bcl-2,Caspase 3,and the cell apoptosis rate increased(P＜0.05).The miR-766-3p upregulation group exhibited a significant decrease in Epac-1 expression(P＜0.05),whereas the levels of miR-766-3p,Bcl-2,Caspase 3,and the rate of cell apoptosis all significantly increased(P＜0.05).In the miR-766-3p downregulated group,the expression of Epac-1 increased(P＜0.05),while the expressions of miR-766-3p,Bcl-2,Capase3 and cell apoptosis rate decreased(P＜0.05).The relative luciferase activity of miR-766-3p WT in the LncRNA H19 group was lower than that in the transfected control group(P＜0.05),and there were binding sites between LncRNA H19 and miR-766-3p.Conclusion:Hypoxia-reoxygenation can lead to a decrease in the expression of LncRNA H19 and Epac-1 in the H9c2 rat myocardial cell line,while the expressions of miR-766-3p,Bcl-2,and Caspase 3 increase,along with a higher cell apoptosis rate.Upregulation of LncRNA H19 can target miR-766-3p and reduce cell apoptosis.
Keywords：hypoxia reoxygenationmyocardial cellsapoptosislong-chain non-coding ribonucleic acid H19micro ribonucleic acid-766-3pexperiment research
Publication Date：2025-08-25
Online Publishing Date：2025-09-10(First online date of this platform, not the publication date of the document)
Pages：7( 2461-2467 )