MiR-421 inhibition protects H9c2 cells against hypoxia/reoxygenation-induced oxidative stress and apoptosis by targeting Sirt3
| Autor: | Yu Liu, Xi-Ming Qian, Qi-Cai He, Jia-Kan Weng |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
SIRT3
Apoptosis Myocardial Reperfusion Injury medicine.disease_cause Transfection 03 medical and health sciences 0302 clinical medicine Sirtuin 3 microRNA medicine Humans Radiology Nuclear Medicine and imaging Myocardial infarction 030304 developmental biology Advanced and Specialized Nursing 0303 health sciences business.industry General Medicine medicine.disease Cell Hypoxia MicroRNAs Oxidative Stress Cancer research Hypoxia reoxygenation Cardiology and Cardiovascular Medicine business Safety Research 030217 neurology & neurosurgery Oxidative stress |
| Zdroj: | Perfusion. 35(3) |
| ISSN: | 1477-111X |
| Popis: | Background: MicroRNAs (miRNAs) are involved in myocardial ischemia-reperfusion injury. miRNA-421 (miR-421) plays a significant role in the initiation of apoptosis and myocardial infarction. However, the molecular regulation of miR-421 in myocardial ischemia-reperfusion injury requires further elucidation. Methods: An in vitro hypoxia/reoxygenation model was established, and the expression levels of miR-421 and Sirtuin-3 (Sirt3) in H9c2 cells were quantified using quantitative real-time polymerase chain reaction. Flow cytometry was employed to measure the effects of miR-421 on myocardial apoptosis induced by hypoxia/reoxygenation. The activity of lactate dehydrogenase and superoxide dismutase and levels of malondialdehyde were measured. The binding sites of miR-421 on Sirt3 were predicted using TargetScan software. A luciferase reporter assay was used to validate the direct targeting of Sirt3 with miR-421. Protein expression levels of Sirt3 and its downstream proteins were evaluated using Western blot analysis. Results: Exposure of H9c2 cells to hypoxia/reoxygenation led to increased apoptosis, levels of malondialdehyde and lactate dehydrogenase, and decreased levels of superoxide dismutase. miR-421 knockdown resulted in decreased apoptosis, levels of lactate dehydrogenase and malondialdehyde, and increased superoxide dismutase levels in H9c2 cells. Hypoxia/reoxygenation significantly decreased the relative expression levels of Sirt3. Down-regulation of Sirt3 resulted from overexpression of miR-421, which directly targeted Sirt3. Knockdown of miR-421 up-regulated Sirt3 expression, inhibited activation of the Jun N-terminal kinase/activator protein 1 pathway and caspase 9/3-dependent cell death. Conclusion: The miR-421-Sirt3-Jun N-terminal kinase/activator protein 1 axis is a novel molecular mechanism that accommodates hypoxia/reoxygenation-induced oxidative stress and apoptosis and provides a new direction for the study and treatment of hypoxia/reoxygenation. |
| Databáze: | OpenAIRE |
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