Molecular Mechanism of LncRNA SNHG14 Regulating miR-506-3p/Beclin-1 Pathway Mediated Myocardial Ischaemia-reperfusion
LI Xiaoling
SUN Jiangbin
LU Zengxue
CHEN Runqi
YANG Tao
Abstract：Objective:To explore the potential mechanism of long non-coding RNA(LncRNAs)small nucleolar RNA host gene 14(SNHG14)regulating the miR-506-3p/Beclin-1 pathway mediated myocardial ischemia-reperfusion injury.Methods:After ligating the left anterior descending(LAD)of the coronary artery to construct rats model of myocardial ischemia-reperfusion(I/R)injury and transfection,they were divided into the sham group,the I/R group,the I/R+si-NC group,and the I/R+si-SNHG14 group.The rats were divided into the sham group,the I/R group,the I/R+NC mimic group,and the I/R+miR-506-3p mimic group.After constructing the hypoxia/reoxygenation(H/R)model and transfection,H9c2 cells were divided into the control group,the H/R group,the H/R+si-NC group,the H/R+si-SNHG14 group.H9c2 cells were divided into the control group,the H/R group,the H/R+NC mimic group,and the H/R+miR-506-3p mimic group.H9c2 cells were divided into the H/R+si-SNHG14+miR-506-3p mimic group,the H/R+si-SNHG14+miR-506-3p inhibitor group,the H/R+si-SNHG14+miR-506-3p inhibitor+si-Beclin-1 group,and the H/R+si-SNHG14+miR-506-3p mimic+ov-Beclin-1 group.H9c2 cells were divided into the co-transfection group of NC mimic and wild-type SNHG14(WT-SNHG14),the co-transfection group of NC mimic and mutant SNHG14(MUT-SNHG14),the co-transfection group of miR-506-3p mimic and WT-SNHG14,and the co-transfection group of miR-506-3p mimic and MUT-SNHG14.H9c2 cells were divided into the co-transfection group of NC mimic and wild-type Beclin-1(WT Beclin-1),the co-transfection group of NC mimic and mutant Beclin-1(MUT Beclin-1),the co-transfection group of miR-506-3p mimic and WT Beclin-1,the co-transfection group of miR-506-3p mimic and MUT Beclin-1.The myocardial infarction area was detected by 2,3,5-triphenyltetrazolium chloride(TTC)staining.The levels of lactate dehydrogenase(LDH)and creatine kinase isoenzyme(CK-MB)were detected by corresponding kits.The cell viability was detected by the CCK-8 assay kit.Cell apoptosis was determined by adopting Annexin V-FITC/PI cell apoptosis detection kit and terminal deoxynucleotide transferase-mediated dUTP gap end labeling assay(TUNEL)staining kit.The levels of SNHG14,miR-506-3p,and Beclin-1 were detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).The expressions of Bax,Bcl-2,and Beclin-1 were detected by Western Blot.The upstream and downstream binding targets of miR-506-3p were predicted by the StarBase software.The targeting relationships between SNHG14 and miR-506-3p,miR-506-3p and Beclin-1 were detected by the luciferase reporter system.Results:In vivo,compared with the sham group,the myocardial infarction area in the I/R group increased(P＜0.001),and the transfection of si-SNHG14 or miR-506-3p mimic reduced the myocardial infarction area(P＜0.001).In vitro,compared with the control group,the apoptosis rate,the protein expression level of Bax,and the gene and protein expression level of Beclin-1 increased,while the cell viability and the protein expression of Bcl-2 decreased in the H/R group(P＜0.001),and the transfection of si-SNHG14 or miR-506-3p mimic reversed the changes in these indicators.After transfection with si-SNHG14,the expression of miR-506-3p increased(P＜0.001).In vivo and in vitro,compared with the sham group or control group,the expressions of SNHG14,CK-MB,and LDH in the I/R group or H/R group increased,the level of miR-506-3p decreased(P＜0.001),and the transfection of si-SNHG14 or miR-506-3p mimic reversed the changes in these indicators.Compared with the co-transfection group of NC mimic and WT-SNHG14,the co-transfection group of NC mimic and MUT-SNHG14,the co-transfection group of miR-506-3p mimic and MUT-SNHG14,the co-transfection group of NC mimic and WT Beclin-1,the co-transfection group of NC mimic and MUT Beclin-1,and the co-transfection group of miR-506-3p mimic and MUT Beclin-1,the dual fluorescence levels in the co-transfection group of miR-506-3p mimic and WT-SNHG14 and the co-transfection group of miR-506-3p mimic and WT Beclin-1 decreased(P＜0.001).Compared with the H/R+si-SNHG14+miR-506-3p inhibitor group,the apoptosis rate,the protein expressions of Bax and Beclin-1,and the levels of LDH and CK-MB decreased,while the cell viability and the protein expression of Bcl-2 increased in the H/R+si-SNHG14+miR-506-3p inhibitor+si-Beclin-1 group.Compared with the H/R+si-SNHG14+miR-506-3p mimic group,the apoptosis rate,the protein expressions of Bax,Beclin-1,LDH and CK-MB increased,while the cell viability and the protein expression of Bcl-2 decreased in the H/R+si-SNHG14+miR-506-3p inhibitor+ov-Beclin-1 group.Conclusion:The LncRNA SNHG14/miR-506-3p/Beclin-1 axis might be a key pathway for the treatment of myocardial ischemia-reperfusion injury.
Keywords：myocardial ischemia-reperfusion injurylong non-coding RNA small nucleolar RNA host gene 14miR-506-3pBeclin-1miceexperimental study
Publication Date：2025-05-25
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：16( 1499-1514 )