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Updated: Dec 27, 2025
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
Downregulation of miR-3568 Protects Against Ischemia/Reperfusion-Induced Cardiac Dysfunction in Rats and Apoptosis in
Xin Li1, Xin Wang1, Yuan-Sheng Liu2
1Department of Cardiovascular Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China.
Abstract:
microRNA-3568 (miR-3568) has been reported to be associated with atherosclerosis. Only few data describe the expression and underlying mechanism of miR-3568 in regulating cardiac ischemia-reperfusion (I/R) injury such as apoptosis. In this study, we therefore sought to investigate the potential function of miR-3568 in simulated I/R-induced apoptosis in H9C2 cardiomyocytes and related signaling pathways involved. Flow cytometry was performed to examine the cell apoptosis. The expression of miR-3568, Survivin, Bcl-2, ERK, JNK, p38, AKT, and STAT3 was measured by western blot and quantitative real-time PCR. The correlation between TRIM62 and p-STAT3 was measured by co-immunoprecipitation and ubiquitination. We found that miR-3568 expression in simulated I/R-induced H9C2 cardiomyocytes was increased in a time-dependent manner. miR-3568 mimic transfection in H9C2 cardiomyocytes significantly enhanced cell apoptosis, decreased the expression of Bcl-2 and Survivin, and activated STAT3 signaling, which were reversed by miR-3568 inhibitor. The direct interaction between miR-3568 and the 3'-untranslated region (UTR) of TRIM62 mRNA was confirmed by dual-luciferase reporter assay. TRIM62 overexpression or AG490, a selective inhibitor of JAK2/STAT3 significantly, significantly inhibited I/R and miR-3568 mimic induced cell apoptosis and STAT3 activation. TRIM62 was found to interact with and induce ubiquitination of p-STAT3. The facilitating role of miR-3568 in I/R injury was also observed in our in vivo rat models. In conclusion, our study suggests that miR-3568 promotes simulated I/R-induced apoptosis in H9C2 cardiomyocytes through targeting TRIM62.
Insights
MicroRNA-3568 (miR-3568) promotes cardiac apoptosis during ischemia-reperfusion (I/R) injury by targeting TRIM62 and activating STAT3 signaling. This finding offers new insights into cardiac I/R injury mechanisms.
Area of Science:
- Cardiovascular Biology
- Molecular Cardiology
- MicroRNA Research
Background:
- MicroRNA-3568 (miR-3568) is linked to atherosclerosis, but its role in cardiac ischemia-reperfusion (I/R) injury and apoptosis is unclear.
- Understanding miR-3568's function in cardiac I/R injury is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the role of miR-3568 in simulated I/R-induced apoptosis in H9C2 cardiomyocytes.
- To elucidate the underlying signaling pathways, including STAT3 activation and TRIM62 interaction.
Main Methods:
- Utilized H9C2 cardiomyocytes and in vivo rat models for I/R injury studies.
- Assessed cell apoptosis via flow cytometry.
- Measured gene and protein expression (miR-3568, Survivin, Bcl-2, ERK, JNK, p38, AKT, STAT3) using quantitative real-time PCR and Western blot.
- Confirmed direct interaction between miR-3568 and TRIM62 mRNA using dual-luciferase reporter assay.
- Investigated TRIM62 and p-STAT3 interaction and ubiquitination via co-immunoprecipitation.
Main Results:
- miR-3568 expression increased time-dependently in simulated I/R conditions.
- miR-3568 mimic enhanced apoptosis, reduced Bcl-2/Survivin, and activated STAT3 signaling in cardiomyocytes.
- miR-3568 directly targets the 3'-UTR of TRIM62 mRNA.
- TRIM62 overexpression or JAK2/STAT3 inhibition reduced I/R and miR-3568 mimic-induced apoptosis and STAT3 activation.
- TRIM62 interacts with and promotes ubiquitination of p-STAT3.
- miR-3568 exacerbated I/R injury in in vivo rat models.
Conclusions:
- miR-3568 promotes simulated I/R-induced apoptosis in H9C2 cardiomyocytes.
- This effect is mediated through the targeting of TRIM62, leading to STAT3 pathway activation.
- Findings highlight miR-3568 as a potential therapeutic target for cardiac I/R injury.
