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Updated: May 7, 2025
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
OGT-mediated O-GlcNAcylation regulates macrophage polarization in heart failure via targeting IRF1
1Department of Cardiovascular Medicine, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, 010050, China.
Insights
Silencing O-linked N-acetylglucosamine (O-GlcNAcylation) enzyme OGT shifts macrophages to an anti-inflammatory state, alleviating heart failure (HF). This suggests O-GlcNAcylation is a potential therapeutic target for HF.
Area of Science:
- Biochemistry
- Immunology
- Cardiovascular Medicine
Background:
- Heart failure (HF) is a global health concern with high mortality.
- Macrophage inflammation significantly contributes to HF pathogenesis.
- O-GlcNAcylation, a post-translational modification, influences various cellular processes.
Purpose of the Study:
- To investigate the role of O-GlcNAcylation in heart failure.
- To determine the impact of O-GlcNAcylation on macrophage polarization in HF.
Main Methods:
- Utilized lipopolysaccharide (LPS) to induce pro-inflammatory macrophages in Raw264.7 cells.
- Generated HF mouse models via transverse aortic constriction (TAC).
- Assessed macrophage polarization using qPCR and flow cytometry after OGT knockdown or IRF1 overexpression; analyzed mechanisms via bioinformatics, co-IP, IP, and western blotting.
Main Results:
- Elevated O-GlcNAcylation and OGT levels were observed in LPS-treated cells.
- OGT knockdown inhibited pro-inflammatory and promoted anti-inflammatory macrophage polarization.
- OGT silencing alleviated TAC-induced cardiac dysfunction and fibrosis by suppressing IRF1 O-GlcNAcylation at Ser283.
Conclusions:
- OGT silencing promotes a shift in macrophage polarization towards an anti-inflammatory phenotype, thereby ameliorating HF.
- This beneficial effect is mediated through the O-GlcNAcylation of IRF1.
- O-GlcNAcylation emerges as a promising therapeutic strategy for heart failure treatment.
Background:
Heart failure (HF) is a syndrome with complex etiology and high mortality in the world. Macrophage-related inflammation is involved in HF development. O-GlcNAcylation is a post-translational modification that affects pathological processes. This study aimed to investigate the role of O-GlcNAcylation in HF, especially its effect on macrophage polarization.
Methods:
Raw264.7 cells were treated with lipopolysaccharide (LPS) to induce pro-inflammatory macrophages. HF mice were generated by transverse aortic constriction (TAC). After knockdown of OGT or overexpressing IRF1, macrophage polarization was evaluated using quantitative real-time polymerase chain reaction and flow cytometry. Underlying mechanism was analyzed using bioinformatic analysis, co-immunoprecipitation (co-IP), IP, and western blotting.
Results:
The results showed that O-GlcNAcylation and OGT levels were high in LPS-treated Raw264.7 cells. OGT knockdown inhibited pro-inflammatory macrophage polarization and promoted anti-inflammatory macrophage polarization caused by LPS, and alleviated TAC-induced cardiac dysfunction and fibrosis. Mechanistically, OGT silence suppressed O-GlcNAcylation of IRF1 at Ser (S)283 site. IRF1 overexpression reversed macrophage polarization modulated by OGT knockdown.
Conclusion:
Silencing of OGT promotes macrophage polarization from pro-inflammatory to anti-inflammatory phenotype to alleviate HF through O-GlcNAcylation of IRF1. The findings suggest that O-GlcNAcylation has the potential to treat HF.
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