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Updated: Mar 7, 2026
Isolation and Flow Cytometric Assessment of Neuroimmune Interactions in a Mini-Stroke Murine Model
Published on: June 20, 2025
Long non-coding RNA SNHG14 promotes microglia activation by regulating miR-145-5p/PLA2G4A in cerebral infarction
Xu Qi1, Ming Shao2, Hongjing Sun1
1Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, China.
Abstract:
Activated microglia cells (MCs) are able to release a large amount of inflammatory cytokines after ischemic stroke, which exacerbates neuron damage. In this study, we explored the functional involvement of long non-coding RNA (lncRNA) SNHG14 and its potential regulatory mechanism in the activation of MCs. The mouse model of middle cerebral artery occlusion (MCAO) and microglia cell model of oxygen/glucose deprivation (OGD) were made. The expression of SNHG14, miR-145-5p and PLA2G4A protein expression was determined by quantitative real time PCR and western blot, respectively. Dual-luciferase assay was used to verify the direct binding of miR-145-5p and PLA2G4A. Flow cytometry was applied to measure neurons' apoptosis. SNHG14 highly expressed in ischemic cerebral tissues and BV-2 cells after OGD treatment. SNHG14 knockdown could remarkably inhibit BV-2 cells activation induced by OGD; while SNHG14 overexpression significantly promoted BV-2 cells activation, showing an increase of TNF-α and NO production and neurons' apoptosis rate. Additionally, SNHG14 knockdown promoted the expression of miR-145-5p and reduced PLA2G4A. Contrarily, SNHG14 overexpression inhibited miR-145-5p expression and increased PLA2G4A. Moreover, miR-145-5p overexpression also reversed the effect of OGD on BV-2 cells activation. Bioinformatics analysis and dual-luciferase assay supported that SNHG14 could bind directly to miR-145-5p and miR-145-5p-binding site was existed on 3'-UTR of PLA2G4A. MiR-145-5p mimic reversed the increase of PLA2G4A and reduced the high levels of TNF-α and NO in BV-2 cells induced by SNHG14 overexpression. SNHG14 increased the expression of PLA2G4A by inhibition of miR-145-5p, which resulted in the activation of MCs in cerebral infarction.
Insights
Long non-coding RNA SNHG14 promotes microglia activation and neuron damage after ischemic stroke by inhibiting miR-145-5p and increasing PLA2G4A. Targeting SNHG14 may offer a therapeutic strategy for cerebral infarction.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Activated microglia cells release inflammatory cytokines, exacerbating neuron damage post-ischemic stroke.
- Long non-coding RNAs (lncRNAs) play roles in cellular processes, but their specific involvement in microglia activation during stroke is under investigation.
Purpose of the Study:
- To investigate the role of lncRNA SNHG14 in microglia activation following ischemic stroke.
- To elucidate the regulatory mechanism of SNHG14, including its interaction with miR-145-5p and PLA2G4A, in microglia activation.
Main Methods:
- Established mouse models of middle cerebral artery occlusion (MCAO) and oxygen/glucose deprivation (OGD) in microglia.
- Quantified gene and protein expression using quantitative real-time PCR and western blot.
- Verified molecular interactions via dual-luciferase assay and flow cytometry for apoptosis analysis.
Main Results:
- SNHG14 was highly expressed in ischemic tissues and OGD-treated microglia.
- SNHG14 knockdown inhibited microglia activation, while overexpression promoted it, increasing inflammatory markers (TNF-α, NO) and neuron apoptosis.
- SNHG14 directly binds to miR-145-5p, inhibiting its expression and leading to increased PLA2G4A, ultimately activating microglia.
Conclusions:
- SNHG14 acts as a crucial regulator in microglia activation post-ischemic stroke.
- The SNHG14/miR-145-5p/PLA2G4A axis contributes to inflammatory responses and neuron damage in cerebral infarction.
- Targeting SNHG14 presents a potential therapeutic avenue for ischemic stroke treatment.
