Abstract
MiR-499a-5p was significantly downregulated in degenerative tissues and correlated with apoptosis. Nonetheless, the biological function of miR-499a-5p in acute ischemic stroke has been still unclear. In this study, we found that the plasma levels of miR-499a-5p were significantly downregulated in 64 ischemic stroke patients and negatively correlated with the National Institutes of Health Stroke Scale score. Then, we constructed cerebral ischemia/reperfusion (I/R) injury in rats after middle cerebral artery occlusion and subsequent reperfusion and oxygen–glucose deprivation and reoxygenation (OGD/R)-treated SH-SY5Y cell model. Transfection with miR-499a-5p mimic was accomplished by intracerebroventricular injection in the in vivo I/R injury model. We further found that miR-499a-5p overexpression decreased infarct volumes and cell apoptosis in the in vivo I/R stroke model using TTC and TUNEL staining. PDCD4 was a direct target of miR-499a-5p by luciferase report assay and Western blotting. Knockdown of PDCD4 reduced the infarct damage and cortical neuron apoptosis caused by I/R injury. MiR-499a-5p exerted neuroprotective roles mainly through inhibiting PDCD4-mediated apoptosis by CCK-8 assay, LDH release assay, and flow cytometry analysis. These findings suggest that miR-499a-5p might represent a novel target that regulates brain injury by inhibiting PDCD4-mediating apoptosis.
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Data Availability
The data in this study are available in this published article.
Abbreviations
- I/R:
-
Ischemia/reperfusion
- OGD/R:
-
Oxygen–glucose deprivation and reoxygenation
- 3′-UTR:
-
3′-Untranslated region
- PDCD4:
-
Programmed cell death 4
- HIF-1α:
-
Hypoxia inducible factor 1 alpha
- MCAO:
-
Middle cerebral artery occlusion
- NIHSS:
-
National Institutes of Health Stroke Scale
- TTC:
-
2,3,5-Triphenyltetrazolium chloride
- TUNEL:
-
TdT-mediated dUTP Nick-end labeling
- DMEM:
-
Dulbecco's modified Eagle's medium
- RT-qPCR:
-
Reverse transcription quantitative PCR
- SDS-PAGE:
-
Sodium dodecyl sulfate–polyacrylamide gel electrophoresis
- LDH:
-
Lactate dehydrogenase
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The use of the human blood samples for research purposes was approved by the Ethics Committee of the 1st Affiliated Hospital, School of Medicine, Zhejiang University (Approval number: ZU349-2; 2017.8.16).
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10571_2021_1085_MOESM1_ESM.tif
Figure 1S Selected four top targets of miR-499a-5p. (A) The potential interaction between miR-499a-5p and putative binding sites in the 3′UTR of four top targets (MRPS35, ARGLU1, PFN2, and HNRNPC). (B) Western blot analysis was used to determine the protein levels of four top targets (MRPS35, ARGLU1, PFN2, and HNRNPC) in SH-SY5Y cells transfected with miR-499a-5p mimic or miR-NC, and then was exposed (TIF 10201 kb)
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Shan, W., Ge, H., Chen, B. et al. Upregulation of miR-499a-5p Decreases Cerebral Ischemia/Reperfusion Injury by Targeting PDCD4. Cell Mol Neurobiol 42, 2157–2170 (2022). https://doi.org/10.1007/s10571-021-01085-4
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DOI: https://doi.org/10.1007/s10571-021-01085-4