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Updated: Jul 6, 2025
Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro
Published on: June 13, 2011
Microglia TREM1-mediated neuroinflammation contributes to central sensitization via the NF-κB pathway in a chronic
Songtang Sun1, Zhenzhen Fan1, Xuejiao Liu1
1Department of Neurology, Lanzhou University Second Hospital, Lanzhou University, Lanzhou, 730000, China.
Background:
Neuroinflammation, mediated by the activation of microglia, contributes to central sensitization, which is associated with the development of chronic migraine (CM). TREM1 receptors amplify the inflammatory response. However, their relationship to CM is unclear. Thus, this study endeavoured to elucidate the exact role of TREM1 in CM.
Methods:
Nitroglycerin (NTG) was repeatedly administered intraperitoneally to establish the CM model. Mechanical and thermal sensitivities were assessed using von Frey filaments and hot plate assays. Using Western blotting, TREM1, NF-κB pathway, NLRP3 inflammasome components, and proinflammatory cytokines were all detected. Immunofluorescence was used to examine the cellular distribution of TREM1 and NLRP3, the number of microglia, immunoreactivity, and morphological changes. We examined the effects of TREM1 antagonists (LR12) and NF-κB inhibitors (PDTC) on pain behaviour, as well as the production of c-fos and CGRP. Additionally, we investigated whether LR12 and PDTC affect the activation of microglia and the NLRP3 inflammasome. We synthesized siRNA and TREM1-overexpressing plasmids to transfect BV2 cells treated with LPS and normal BV2 cells and treated TREM1-overexpressing BV2 cells with PDTC. The NF-κB pathway, NLRP3 inflammasome components, and proinflammatory cytokines were quantified using Western blotting.
Results:
Following NTG administration, the expression of TREM1 was significantly upregulated and exclusively localized in microglia in the TNC, and was well co-localized with NLRP3. Furthermore, activation of the classical NF-κB pathway was observed. Pre-treatment with LR12 and PDTC effectively attenuated mechanical hypersensitivity, suppressed the expression of c-fos and CGRP, and inhibited NF-κB activity in CM mice. Additionally, inhibition of TREM1 and NF-κB activity mitigated NTG-induced microglia and NLRP3 activation, as well as proinflammatory cytokines production. In vitro, knockdown of TREM1 resulted in attenuated activation of the NF-κB pathway following lipopolysaccharide (LPS) treatment and reduced expression of NLRP3 inflammasome components as well as proinflammatory cytokines. After TREM1 overexpression, the NF-κB pathway was activated, NLRP3 inflammasome components and proinflammatory cytokines were upregulated, and PDTC reversed this phenomenon.
Conclusions:
Our findings suggest that TREM1 regulates microglia and NLRP3 activation via the NF-κB pathway, thereby contributing to central sensitization and implicating its involvement in chronic migraine pathogenesis.
Insights
Triggering receptor expressed on myeloid cells 1 (TREM1) exacerbates chronic migraine by activating microglia and the NLRP3 inflammasome via the NF-κB pathway. Inhibiting TREM1 reduces neuroinflammation and pain hypersensitivity.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Neuroinflammation, driven by microglial activation, is crucial for central sensitization in chronic migraine (CM).
- TREM1 receptors amplify inflammatory responses, but their role in CM remains unclear.
Purpose of the Study:
- To investigate the role of TREM1 in the pathogenesis of chronic migraine.
- To elucidate the mechanisms by which TREM1 influences neuroinflammation and pain in CM.
Main Methods:
- Established a chronic migraine model in mice using nitroglycerin (NTG) administration.
- Assessed pain behaviors, neuroinflammation markers (TREM1, NF-κB, NLRP3), and pro-inflammatory cytokines using Western blotting and immunofluorescence.
- Investigated the effects of TREM1 antagonists and NF-κB inhibitors in vivo and in vitro using cell transfection and LPS stimulation.
Main Results:
- NTG administration upregulated TREM1 in microglia within the trigeminal nucleus caudalis (TNC), co-localizing with NLRP3 and activating the NF-κB pathway.
- TREM1 and NF-κB inhibition attenuated pain hypersensitivity, reduced c-fos and CGRP expression, and suppressed microglial and NLRP3 inflammasome activation.
- In vitro, TREM1 knockdown decreased NF-κB activation, NLRP3 components, and cytokine production; TREM1 overexpression increased these markers, which were reversed by NF-κB inhibition.
Conclusions:
- TREM1 plays a significant role in chronic migraine pathogenesis by regulating microglial and NLRP3 inflammasome activation through the NF-κB pathway.
- Targeting TREM1 offers a potential therapeutic strategy for chronic migraine by mitigating central sensitization and neuroinflammation.
