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Published on: January 30, 2014
The E3 Ubiquitin Ligase c-Cbl Inhibits Microglia-Mediated CNS Inflammation by Regulating PI3K/Akt/NF-κB Pathway
Lin Dong1,2,3, Yu-Zhen Li4, Hai-Ting An1,2,3
1Department of Neurobiology, Capital Medical University, Beijing, China.
Background:
Microglia-mediated inflammation may play an important role in the pathophysiology progression of neurodegenerative diseases, such as Parkinson's disease (PD), but the molecular mechanisms are poorly understood.
Aims:
This study sought to determine whether E3 ubiquitin ligase c-Cbl plays a role in the brain inflammation and to explore the relevant molecular mechanism.
Methods:
After BV2 microglial cells and c-Cbl-deficient mice were treated with lipopolysaccharide (LPS), neuroinflammation and microglial activation were evaluated by immunohistochemistry, ELISA and Western blot. We further investigated the possible mechanism of c-Cbl in regulating microglial activation.
Results:
Here, we showed that the E3 ubiquitin ligase c-Cbl had high expression in brain tissues including substantia nigra pars compacta (SNc), striatum and hippocampus, and it was abundantly expressed in microglia. Systemic LPS administration resulted in more severe microglial activation in CNS and increased expression of brain proinflammatory factors (TNF-α, IL-6, IL-1β and MCP-1) in c-Cbl knockout mice than wild type mice (WT). Downregulation of c-Cbl expression with c-Cbl siRNA in BV-2 microglial cells demonstrated a more robust increase in the proinflammatory factors release and NF-κB p65 nuclear translocation than that in control siRNA. Interestingly, Akt phosphorylation induced by LPS was also significantly augmented after c-Cbl knockdown. Moreover, blockade of PI3K/Akt activation by LY294002 significantly reduced inflammation response and NF-κB p65 nuclear translocation.
Conclusion:
In sum, c-Cbl inhibits expression of LPS-stimulated proinflammatory cytokines and chemokines in microglia. We demonstrate an unprecedented role for c-Cbl in microglia-mediated neuroinflammation involving PI3K/Akt/NF-κB pathway.
Insights
The E3 ubiquitin ligase c-Cbl inhibits microglia-mediated neuroinflammation. Loss of c-Cbl exacerbates brain inflammation via the PI3K/Akt/NF-κB pathway, revealing a novel therapeutic target for neurodegenerative diseases.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Microglia-mediated inflammation is implicated in neurodegenerative diseases like Parkinson's disease (PD).
- The precise molecular mechanisms driving this inflammation remain poorly understood.
- E3 ubiquitin ligase c-Cbl's role in brain inflammation is yet to be elucidated.
Purpose of the Study:
- To investigate the role of E3 ubiquitin ligase c-Cbl in brain inflammation.
- To explore the molecular mechanisms underlying c-Cbl's involvement in neuroinflammation.
- To determine if c-Cbl influences microglial activation and inflammatory responses.
Main Methods:
- Assessed neuroinflammation and microglial activation in BV2 microglial cells and c-Cbl-deficient mice treated with lipopolysaccharide (LPS).
- Utilized immunohistochemistry, ELISA, and Western blot to evaluate inflammatory markers and protein expression.
- Investigated the impact of c-Cbl knockdown and PI3K/Akt pathway inhibition on inflammatory signaling.
Main Results:
- c-Cbl is highly expressed in microglia within key brain regions.
- c-Cbl knockout mice exhibited exacerbated neuroinflammation and increased pro-inflammatory factors post-LPS treatment.
- Downregulation of c-Cbl enhanced LPS-induced pro-inflammatory cytokine release and NF-κB activation.
- c-Cbl knockdown augmented Akt phosphorylation, which was reversed by PI3K/Akt pathway blockade.
Conclusions:
- c-Cbl acts as an inhibitor of LPS-stimulated pro-inflammatory cytokine and chemokine expression in microglia.
- This study reveals a novel role for c-Cbl in regulating microglia-mediated neuroinflammation.
- The mechanism involves the PI3K/Akt/NF-κB signaling pathway, suggesting c-Cbl as a potential therapeutic target.
