Study on mechanism of LncRNA NEAT1 on improving myocardial injury in rats with myocardial infarction through miR-136/ERK1/2 axis
10.3969/j.issn.1000-484X.2025.01.012
- VernacularTitle:LncRNA NEAT1通过miR-136/ERK1/2轴对改善心肌梗死大鼠心肌损伤的机制研究
- Author:
Bao YIN
1
;
Xiangyu TAN
Author Information
1. 淄博市中医医院,淄博 255300
- Publication Type:Journal Article
- Keywords:
Long non-coding RNA nuclear-enrichment transcript 1;
miR-136;
Myocardial infarction;
Myocardial injury
- From:
Chinese Journal of Immunology
2025;41(1):75-84
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of long non-coding RNA nuclear-enriched transcript 1(LncRNA NEAT1)on improving myocardial injury in rats with myocardial infarction(MI)mechanism through miR-136/extracellular signal-regulated kinase 1/2(ERK1/2)axis.Methods:Using the random number table method,120 male SD rats were divided into 6 groups(n=20):sham group,myocardial infarction group(MI group),control interference group(sh-NC group),NEAT1 interference group(sh-NEAT1 group),NEAT1 interference+miR-136 control inhibition group(sh-NEAT1+antagomiR-NC group)and NEAT1 interference+miR-136 suppression group(sh-NEAT1+antagomiR-136 group).Seven days before MI operation,100 μl of corresponding adenovirus vector was injected into the myocardium.Rats in each group were ligated the left anterior descending vessel to establish MI model.In sham group,only the left chest was opened and the heart was exposed without ligation.H9C2 cardiomyocyte were divided into:control group,vehicle group,sh-NC group,sh-NEAT1 group,sh-NEAT1+antagomiR-NC group and sh-NEAT1+antagomiR-136 group.Car-diomyocyte were transfected with corresponding NEAT1 interference vector and negative control,miR-136 inhibitor and negative con-trol.StarBase prediction and dual luciferase report experiment verify the targeted regulation of LncRNA NEAT1 on miR-136;myocar-dial infarct size was measured by TTC staining,and myocardial histopathological changes were observed by HE staining;Echocardiog-raphy was used to detect rat cardiac function;detection of apoptosis in rat myocardial tissue and H9C2 cells by flow cytometry.ELISA method was used to detect serum oxidative stress,corresponding indexes of myocardial enzymes and the contents of inflammatory fac-tors in myocardial tissue and H9C2 cells.CCK8 and EdU staining were used to detect the proliferation ability of H9C2 cells.RT-qPCR was used to detect the expressions of miR-136,SIRT1 mRNA in myocardial tissue and H9C2 cells.Western blot was used to detect the expressions of ERK1/2,p-ERK1/2,B-cell lymphoma-2(Bcl-2),Bcl2-related X protein(Bax)protein in rat myocardial tissue and H9C2 cells.Results:The expression of LncRNA NEAT1 was up-regulated in injured myocardial tissues and cells,and the expression of miR-136 was down-regulated(P<0.05).At the same time,NEAT1 targets and regulates the level of miR-136.Inhibition of NEAT1 expression could increase left ventricular ejection fraction(LVEF),left ventricular fraction shortening(LVFS),Bcl-2 level,and de-crease the activities of creatine kinase(CK),lactate dehydrogenase(LDH),and aspartame Acid transaminase(AST),superoxide dismutase(SOD),reduce Bax,troponin Ⅰ(cTnⅠ),malondialdehyde(MDA),TNF-α,IL-1β,IL-6,ERK1/2/p-ERK1/2 levels(P<0.05),promote the proliferation of myocardial cells,inhibit cell apoptosis,reduce the area of myocardial infarction,inhibit oxida-tive stress and inflammation,and improve myocardial damage Inhibiting miR-136 can save the above effects.Conclusion:The expres-sion of LncRNA NEAT1 is up-regulated and the expression of miR-136 is down-regulated in injured myocardial tissue and cells.Inhibi-tion of LncRNA NEAT1 expression inhibits apoptosis,oxidative stress and inflammation through miR-136/ERK1/2 axis,thus improving myocardial injury.
