Related Experiment Video
Updated: Nov 29, 2025
Protection of H9c2 Myocardial Cells from Oxidative Stress by Crocetin via PINK1/Parkin Pathway-Mediated Mitophagy
Published on: May 26, 2023
Inhibition of microRNA-184 reduces H2O2-mediated cardiomyocyte injury via targeting FBXO28
1Department of Cardiology, People's Hospital of Rizhao, Rizhao, China. 576831670@qq.com.
Objective:
Cardiovascular disease, especially coronary heart disease, is one of the diseases with the highest mortality. A large number of studies have found that microRNAs (miRNAs) are closely related to the occurrence and development of myocardial ischemia. This article mainly focused on the regulation of miR-184 on oxidative stress, inflammation, and apoptosis in myocardial infarction (MI).
Materials And Methods:
MiR-184 inhibitor or negative control (NC) were transfected into H9c2 cells. Then, H9c2 cells were treated with H2O2 to construct a cardiomyocyte injury model. H9c2 cells were divided into 4 groups: control group, H22O2 treatment group, H2O2 + NC group, and H2O2 + miR-184 inhibitor group. The oxidative stress of H9c2 cells was observed by the expression levels of SOD, ROS, and MDA in each group. The inflammatory response of H9c2 cells was reflected by the expression of TNF-α, IL-6, and IL-1β detected by ELISA kits. Western blot was used to detect the expression of cleaved Caspase-3, Bcl-2, Bax and F-box protein 28 (FBXO28). Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was utilized to detect miR-184 expression. TdT-mediated dUTP Nick-End Labeling (TUNEL) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay were used to observe the apoptosis and cell viability. The Luciferase reporter experiment was used to prove whether miR-184 could target FBXO28.
Results:
MiR-184 expression was significantly increased in H2O2-induced H9c2 cell injury model. After H9c2 cells were transfected with miR-184 inhibitor to silence miR-184, the levels of ROS and MDA were markedly reduced, while the expression of SOD was greatly increased. At the same time, the expression of inflammatory factors was greatly reduced. Silencing miR-184 also increased Bcl-2 expression, and reduced the expression of cleaved Caspase-3 and Bax. In addition, compared with the H2O2 + NC group, the number of TUNEL positive cells in the H2O2 + miR-184 inhibitor group was also significantly reduced, and the cell viability was remarkably increased. The Luciferase reporter experiment proved that FBXO28 is a target gene of miR-184.
Conclusions:
MiR-184 expression was increased in H2O2-treated H9c2 cells. Inhibition of miR-184 markedly inhibited oxidative stress and inflammation in cardiomyocytes, thereby inhibiting cardiomyocyte apoptosis, through the regulation of FBXO28.
Insights
Inhibiting microRNA-184 (miR-184) reduces oxidative stress and inflammation in myocardial infarction models. This protective effect is mediated by regulating the F-box protein 28 (FBXO28), thereby inhibiting cardiomyocyte apoptosis.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- MicroRNA Research
Background:
- Cardiovascular diseases, particularly coronary heart disease, are leading causes of mortality.
- MicroRNAs (miRNAs) play a critical role in the pathogenesis of myocardial ischemia.
- Understanding miRNA regulation in myocardial infarction (MI) is crucial for therapeutic development.
Purpose of the Study:
- To investigate the regulatory role of miR-184 in oxidative stress, inflammation, and apoptosis in myocardial infarction.
- To elucidate the underlying molecular mechanisms involving F-box protein 28 (FBXO28).
Main Methods:
- Constructed a cardiomyocyte injury model using H2O2-treated H9c2 cells.
- Utilized miR-184 inhibitors to silence miR-184 expression.
- Assessed oxidative stress (SOD, ROS, MDA), inflammation (TNF-α, IL-6, IL-1β), apoptosis (cleaved Caspase-3, Bcl-2, Bax, TUNEL), and cell viability (MTT).
- Confirmed the targeting of FBXO28 by miR-184 using a luciferase reporter assay.
Main Results:
- MiR-184 expression was upregulated in the H2O2-induced injury model.
- Inhibition of miR-184 significantly reduced oxidative stress and inflammatory markers.
- Silencing miR-184 decreased apoptosis and increased cell viability.
- FBXO28 was identified as a direct target gene of miR-184.
Conclusions:
- MiR-184 is upregulated in myocardial injury and contributes to oxidative stress and inflammation.
- Inhibiting miR-184 exerts protective effects by reducing oxidative stress, inflammation, and apoptosis in cardiomyocytes.
- The miR-184/FBXO28 axis is a key regulator of cardiomyocyte injury and apoptosis.
