lncRNA MEG3 inhibits hypoxia/reoxygenation-induced cardiomyocyte apoptosis by regulating the miR-202-5p/STAT3 axis
10.12007/j.issn.0258-4646.2025.08.011
- VernacularTitle:lncRNA MEG3通过调节miR-202-5p/STAT3轴抑制缺氧/复氧诱导的心肌细胞凋亡
- Author:
Zhiyang WANG
1
;
Zhandong LIU
;
Limei PIAO
;
Chunzi JIN
;
Wenhu XU
Author Information
1. 延边大学附属医院心血管内科,吉林延吉 133000
- Publication Type:Journal Article
- Keywords:
long non-coding RNA maternal expression gene 3;
miR-202-5p/signal transducer and activator of transcription 3 axis;
hypoxia/reoxygenation;
cardiomyocyte
- From:
Journal of China Medical University
2025;54(8):733-739
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impact of long non-coding RNA maternal expression gene 3(lncRNA MEG3)on hypoxia/reoxygenation(H/R)-induced cardiomyocyte apoptosis by modulating the miR-202-5p/signal transducer and activator of transcription 3(STAT3)axis.Methods An H/R cell model was constructed and randomly separated into four groups:sh-NC(transfected with NC shRNA),sh-MEG3(transfected with lncRNA MEG3 shRNA),miR-NC(transfected with lncRNA MEG3 shRNA and NC miR),and in-miR-202-5p(transfected with lncRNA MEG3 shRNA and miR-202-5p inhibitor).In addition,the H/R group(nontransfected H/R model cells)and the AC 16 group(normal AC 16 cells)were set.Quantitative real-time PCR method was used to analyze the expression of lncRNA MEG3,miR-202-5p,and STAT3 in cells from each group.CCK-8 method was used to analyze cell viability.Flow cytometry was used to analyze apoptosis.Enzyme-linked immunosorbent,colorimetric,and probe assays were applied to detect the levels of inter-leukin-6(IL-6),tumor necrosis factor α(TNF-α),malondialdehyde(MD A),glutathione peroxidases(GSH-Px),and reactive oxygen spe-cies(ROS).Western blotting was carried out to examine the expression of STAT3,Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),caspase-3,and caspase-9.A dual luciferase assay was used to analyze the relationship between lncRNA MEG3 and miR-202-5p,as well as the relationship between miR-202-5p and STAT3.Results Compared with that in the AC 16 group,the expression of lncRNA MEG3 and STAT3 in cells in the H/R group increased,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the expression of lncRNA MEG3 and STAT3 decreased in the sh-MEG3 group,while the expression of miR-202-5p increased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the expression of lncRNA MEG3 and STAT3 increased in the in-miR-202-5p group,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the AC 16 group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the H/R group.In contrast,the cell viability,clone count,levels of superoxide dismutase(SOD)and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 increased in the sh-MEG3 group.In contrast,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 decreased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the in-miR-202-5p group.In contrast,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).There were multiple binding sites between lncRNA MEG3 and miR-202-5p,and between miR-202-5p and STAT3.The luciferase activity was lower in the WT-MEG3+miR-202-5p group than in the WT-MEG3+miR-NC group(P＜0.05).The luciferase activity was lower in the WT-STAT3+miR-202-5p group than in the WT-STAT3+miR-NC group(P＜0.05).Conclusion Knocking down lncRNA MEG3 can downregulate STAT3 by negatively regulating miR-202-5p,inhibiting H/R-induced cardiomyocyte apoptosis.
