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Updated: Oct 7, 2025
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
MicroRNA-214-3p facilitates M2 macrophage polarization by targeting GSK3B
Ling-Yan Peng1, Bi-Bao Li2, Ke-Bin Deng1
1Department of Otorhinolaryngology, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, China.
Abstract:
Allergic rhinitis (AR) is a chronic inflammatory disease of the nasal mucosa. M2 macrophage polarization can reduce inflammation and repair tissue injury during AR development. Studies have substantiated the involvement of miRNAs in AR pathogenesis. Herein, the molecular mechanism of miR-214-3p in AR development was explored. To mimic the AR environment, ovalbumin (OVA) was used to treat macrophages. MiR-214-3p and glycogen synthase kinase 3 beta (GSK3B) expression in nasal mucus tissues and macrophages was assessed by RT-qPCR. The M2 phenotypic signature of CD206 in macrophages was assessed by flow cytometry. The protein expression of GSK3B and M2 macrophage markers (ARG-1 and IL-10) was evaluated by western blotting. The correlation between miR-214-3p and GSK3B was validated by a luciferase reporter assay. We found that miR-214-3p was overexpressed in macrophages and nasal mucus tissues from AR patients. MiR-214-3p facilitated M2 polarization of macrophages upon OVA stimulation. Mechanistically, miR-214-3p targeted the GSK3B 3' untranslated region in macrophages. In addition, GSK3B was downregulated in macrophages and nasal mucus tissues from AR patients. In rescue assays, GSK3B downregulation reversed the inhibitory effects of miR-214-3p silencing on M2 polarization of macrophages treated with OVA. Overall, miR-214-3p facilitates M2 macrophage polarization by targeting GSK3B.
Insights
MicroRNA-214-3p promotes M2 macrophage polarization, crucial for allergic rhinitis (AR) inflammation resolution, by targeting GSK3B. This finding offers a new therapeutic target for AR treatment.
Area of Science:
- Immunology
- Molecular Biology
- Allergy Research
Background:
- Allergic rhinitis (AR) is a chronic nasal inflammatory condition.
- M2 macrophage polarization is key to reducing inflammation and repairing tissue damage in AR.
- MicroRNAs (miRNAs) are implicated in the pathogenesis of AR.
Purpose of the Study:
- To investigate the molecular mechanism of miR-214-3p in the development of allergic rhinitis.
- To explore the role of miR-214-3p in M2 macrophage polarization within an AR context.
Main Methods:
- Macrophages were stimulated with ovalbumin (OVA) to mimic the AR environment.
- Expression levels of miR-214-3p and GSK3B were measured using RT-qPCR.
- M2 macrophage markers (CD206, ARG-1, IL-10) and GSK3B protein were assessed via flow cytometry and western blotting.
- A luciferase reporter assay validated the interaction between miR-214-3p and GSK3B.
Main Results:
- MiR-214-3p was found to be overexpressed in macrophages and nasal tissues of AR patients.
- MiR-214-3p enhanced M2 polarization of macrophages stimulated with OVA.
- MiR-214-3p directly targeted GSK3B, which was downregulated in AR patient samples.
- Silencing miR-214-3p inhibited M2 polarization, an effect reversed by GSK3B downregulation.
Conclusions:
- MiR-214-3p plays a significant role in facilitating M2 macrophage polarization in allergic rhinitis.
- The mechanism involves miR-214-3p targeting and downregulating GSK3B.
- This miR-214-3p/GSK3B pathway represents a potential therapeutic target for managing allergic rhinitis.
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