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miR-497-5p promoted neuronal injury in ischemic stroke by inhibiting the BDNF/TrkB/PI3K/Akt pathway
Chunyan Gong1, Xiaona He1, Guiliang Li1
1Department of Rehabilitation Medicine, Qujing No. 1 Hospital, Qujing, Yunnan, China.
Abstract:
The aim of this study was to investigate the molecular mechanism by which miR-497-5p regulates neuronal injury after ischemic stroke through the BDNF/TrkB/Akt signaling pathway. PC12 cells were used to construct a stroke injury model by oxygen-glucose deprivation/reoxygenation (OGD/R). The expression level of miR-497-5p was measured by RT-qPCR. CCK-8 kit was used to detect cell viability. Cell apoptosis and reactive oxygen species (ROS) were detected by flow cytometry. MDA and SOD detection kits were used to detect MDA content and SOD activity. A double luciferase reporter system was used to verify the targeting relationship between miR-497-5p and BDNF. The expression of BDNF, TrkB, p-TrkB, Akt and p-Akt was detected by Western blot. We have found that miR-497-5p expression was inhibited after treatment with OGD/R. Simultaneously, cell apoptosis, MDA content and ROS were upregulated, while cell viability and SOD were significantly decreased in PC12 cells. The effects of OGD/R on PC12 cells were reversed with the downregulation of miR-497-5p. A double luciferase reporter assay demonstrated that miR-497-5p negatively targets BDNF. BDNF inhibited cell apoptosis and oxidative stress injury in PC12 cells. These findings suggest that miR-497-5p aggravates neuronal injury in experimental model of ischemic stroke by inhibiting the BDNF/TrkB/PI3K/Akt signaling pathway.
Insights
MicroRNA-497-5p worsens brain injury after stroke by blocking the BDNF/TrkB/Akt pathway. Inhibiting miR-497-5p may protect neurons from ischemic stroke damage.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Ischemic stroke causes significant neuronal injury.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes, including neuronal survival and death.
- The specific role of miR-497-5p in ischemic stroke remains to be fully elucidated.
Purpose of the Study:
- To investigate the molecular mechanism of miR-497-5p in regulating neuronal injury following ischemic stroke.
- To explore the involvement of the Brain-Derived Neurotrophic Factor (BDNF)/Tropomyosin receptor kinase B (TrkB)/Akt signaling pathway in this process.
Main Methods:
- Established an in vitro ischemic stroke model using PC12 cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R).
- Quantified miR-497-5p expression using RT-qPCR.
- Assessed cell viability (CCK-8), apoptosis, and reactive oxygen species (ROS) via flow cytometry.
- Measured oxidative stress markers (MDA and SOD).
- Verified the interaction between miR-497-5p and BDNF using a dual-luciferase reporter assay.
- Analyzed protein expression levels of BDNF, TrkB, p-TrkB, Akt, and p-Akt through Western blotting.
Main Results:
- OGD/R treatment significantly decreased miR-497-5p expression in PC12 cells.
- Downregulation of miR-497-5p exacerbated OGD/R-induced neuronal injury, increasing apoptosis and ROS, while decreasing cell viability and SOD activity.
- miR-497-5p was confirmed to directly target and inhibit BDNF expression.
- Overexpression of BDNF attenuated OGD/R-induced apoptosis and oxidative stress.
Conclusions:
- miR-497-5p aggravates neuronal injury in an experimental ischemic stroke model.
- This detrimental effect is mediated by the inhibition of the BDNF/TrkB/Akt signaling pathway.
- Targeting miR-497-5p may represent a potential therapeutic strategy for ischemic stroke.
