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Updated: Jul 9, 2025
Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery
Published on: August 16, 2019
Effect of miR-182-5p on apoptosis in myocardial infarction
Nan Niu1, Huangtai Miao2, Hongmei Ren3
1College of Physics and Optoelectronic Engineering, Canghai Campus of Shenzhen University, Shenzhen, Guangdong, 518060, PR China.
Objective:
This study aimed to delineate the diagnostic significance of miR-182-5p by investigating its influence on myocardial apoptosis and function, employing both in vivo and in vitro myocardial infarction models.
Methods:
A rat myocardial infarction model was established. Myocardial infarction area was detected using the 2,3,5-chlorotriphenyltetrazolium (TTC) method, myocardial enzyme spectrums were measured using enzyme-linked immunosorbent assay (ELISA), myocardial structure was detected by hematoxylin and eosin (HE) staining, myocardial apoptosis was detected using the TUNEL method, and expression levels of miR-182-5p and apoptosis-related molecules were detected using real-time fluorescence quantitative PCR (qPCR) and Western blot. miR-182-5p mimics and inhibitor were transfected into rat H9C2 cardiomyocytes and mouse HL-1 cardiomyocytes to establish a hypoxia model. Cardiomyocyte viability was detected using the CCK-8 method, expression levels of apoptosis-related indicators were detected using Western blot, and caspase-3/7 activity was detected using a caspase-3/7 activity detection kit. AAV9 adeno-associated virus was used to construct an miR-182-5p overexpression virus, which was injected into mice through the tail vein to create a mouse myocardial infarction model. TTC, ELISA, HE staining, echocardiography, real-time fluorescence qPCR, and Western blot methods were used to detect the effects of AAV9-miR-182-5p on myocardial injury, myocardial function, and myocardial apoptosis levels in myocardial infarction.
Results:
The rat model displayed reduced miR-182-5p expression concurrent with an increase in apoptosis. The in vitro H9C2 and HL-1 hypoxia models revealed that miR-182-5p augmented the hypoxia-induced decrease in myocardial cell viability, suppressed Bcl-2 expression, and increased Bax, Bnip3, and caspase-3/7 activity levels. The injection of AAV9-miR-182-5p significantly exacerbated myocardial tissue damage, impaired myocardial function, and enhanced apoptosis.
Conclusion:
miR-182-5p escalates myocardial injury during myocardial infarction by fostering apoptosis. Interventions that aim to reduce miR-182-5p levels might be crucial in halting the progression of myocardial infarction.
Insights
MicroRNA-182-5p (miR-182-5p) worsens myocardial infarction by increasing heart cell apoptosis. Reducing miR-182-5p levels may be a key strategy to treat myocardial infarction.
Area of Science:
- Cardiovascular Biology
- Molecular Cardiology
- Biomarker Discovery
Background:
- Myocardial infarction (MI) is a leading cause of mortality worldwide.
- Understanding the molecular mechanisms underlying MI pathogenesis is crucial for developing effective treatments.
- MicroRNAs (miRNAs) have emerged as critical regulators of cardiovascular diseases, including MI.
Purpose of the Study:
- To investigate the role of miR-182-5p in myocardial apoptosis and function.
- To explore the diagnostic significance of miR-182-5p in myocardial infarction.
- To evaluate the therapeutic potential of targeting miR-182-5p in MI.
Main Methods:
- Established in vivo (rat and mouse) and in vitro (cardiomyocyte) models of myocardial infarction and hypoxia.
- Utilized techniques including TTC staining, ELISA, HE staining, TUNEL assay, echocardiography, qPCR, Western blot, and CCK-8 assay.
- Employed miR-182-5p mimics, inhibitors, and adeno-associated virus (AAV9) for gene manipulation.
Main Results:
- Reduced miR-182-5p expression correlated with increased apoptosis in a rat MI model.
- In vitro, miR-182-5p exacerbated hypoxia-induced cardiomyocyte death, suppressed Bcl-2, and increased Bax, Bnip3, and caspase-3/7 activity.
- AAV9-mediated miR-182-5p overexpression worsened myocardial damage, impaired function, and enhanced apoptosis in mice.
Conclusions:
- miR-182-5p promotes myocardial injury and apoptosis in the context of myocardial infarction.
- Targeting miR-182-5p presents a potential therapeutic strategy for mitigating MI progression.
