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miR-542-3p-Targeted PDE4D Regulates cAMP/PKA Signaling Pathway and Improves Cardiomyocyte Injury
1Affiliated Hospital of Jiaxing University, The First Hospital of Jiaxing, Jiaxing 314000, Zhejiang, China.
Objective:
To investigate the protective effect of miR-542-3p on cardiomyocyte injury and related mechanisms.
Methods:
A cardiomyocyte hypoxia/reoxygenation model was established. The expression levels of miR-542-3p and PDE4D were detected using qRT-PCR; the luciferase reporter assay system was used to detect the targeting relationship between miR-542-3p and PDE4D; overexpressing miR-542-3p was transfected into cardiomyocytes, and ROS release was detected by immunofluorescence while cellular apoptosis was detected by TUNEL; and the western blot assay was applied to detect the expression of PDE4D, phosphorylated protein kinase A (p-PKA), and phosphorylated cyclic adenosine monophosphate (cAMP) response element-binding protein (p-CREB).
Results:
Compared with the control group, the miR-542-3p expression level was decreased and the PDE4D expression level was increased in the cardiomyocyte hypoxia/reoxygenation model group. The dual-luciferase reporter assay system confirmed that miR-542-3p could target and regulate PDE4D; the transfection with cardiomyocytes using the overexpressing miR-542-3p could downregulate PDE4D expression, attenuate ROS release during cardiomyocyte injury, and reduce cellular apoptosis rate, while upregulating the expression of p-PKA and p-CREB.
Conclusion:
The miR-542-3p can negatively regulate PDE4D protein expression and attenuate cardiomyocyte injury through a mechanism related to the activation of the cAMP/PKA signaling pathway.
Insights
MicroRNA-542-3p protects heart cells from injury by targeting PDE4D and activating the cAMP/PKA pathway. This finding offers a potential therapeutic target for cardiomyocyte damage.
Area of Science:
- Cardiology
- Molecular Biology
- Biochemistry
Background:
- Cardiomyocyte injury is a significant contributor to heart disease.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes, including cell survival and death.
- Understanding the specific roles of miRNAs in cardiac protection is essential for developing novel therapeutic strategies.
Purpose of the Study:
- To investigate the protective effect of miR-542-3p against cardiomyocyte injury.
- To elucidate the underlying molecular mechanisms, focusing on its interaction with PDE4D and the cAMP/PKA signaling pathway.
Main Methods:
- Establishment of a cardiomyocyte hypoxia/reoxygenation injury model.
- Quantification of miR-542-3p and PDE4D expression using qRT-PCR.
- Luciferase reporter assay to confirm the targeting relationship between miR-542-3p and PDE4D.
- Assessment of reactive oxygen species (ROS) release, cellular apoptosis, and protein expression (PDE4D, p-PKA, p-CREB) via immunofluorescence, TUNEL, and Western blot assays.
Main Results:
- miR-542-3p expression was decreased, while PDE4D expression was increased in the hypoxia/reoxygenation model.
- miR-542-3p directly targeted and downregulated PDE4D.
- Overexpression of miR-542-3p reduced ROS release and apoptosis, and increased p-PKA and p-CREB levels in cardiomyocytes.
Conclusions:
- miR-542-3p exerts a protective effect on cardiomyocytes against hypoxia/reoxygenation injury.
- This protection is mediated by the negative regulation of PDE4D and subsequent activation of the cAMP/PKA signaling pathway.
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