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Published on: March 26, 2019
Targeting the shift from M1 to M2 macrophages in experimental autoimmune encephalomyelitis mice treated with fasudil
Chunyun Liu1, Yanhua Li, Jiezhong Yu
1Institute of Brain Science, Department of Neurology, Medical School, Shanxi Datong University, Datong, China.
Abstract:
We observed the therapeutic effect of Fasudil and explored its mechanisms in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Fasudil, a selective Rho kinase (ROCK) inhibitor, was injected intraperitoneally at 40 mg/kg/d in early and late stages of EAE induction. Fasudil ameliorated the clinical severity of EAE at different stages, and decreased the expression of ROCK-II in spleen, accompanied by an improvement in demyelination and inhibition of inflammatory cells. Fasudil mainly inhibited CD4(+)IL-17(+) T cells in early treatment, but also elevated CD4(+)IL-10(+) regulatory T cells and IL-10 production in late treatment. The treatment of Fasudil shifted inflammatory M1 to anti-inflammatory M2 macrophages in both early and late treatment, being shown by inhibiting CD16/32, iNOS, IL-12, TLR4 and CD40 and increasing CD206, Arg-1, IL-10 and CD14 in spleen. By using Western blot and immunohistochemistry, iNOS and Arg-1, as two most specific markers for M1 and M2, was inhibited or induced in splenic macrophages and spinal cords of EAE mice treated with Fasudil. In vitro experiments also indicate that Fasudil shifts M1 to M2 phenotype, which does not require the participation or auxiliary of other cells. The polarization of M2 macrophages was associated with the decrease of inflammatory cytokine IL-1β, TNF-α and MCP-1. These results demonstrate that Fasudil has therapeutic potential in EAE possibly through inducing the polarization of M2 macrophages and inhibiting inflammatory responses.
Insights
Fasudil, a ROCK inhibitor, effectively treats experimental autoimmune encephalomyelitis (EAE), a multiple sclerosis model. It reduces inflammation by shifting macrophages to an anti-inflammatory M2 phenotype, offering therapeutic potential for neurodegenerative diseases.
Area of Science:
- Neuroimmunology
- Pharmacology
Background:
- Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system.
- Experimental autoimmune encephalomyelitis (EAE) serves as a key animal model for studying MS pathogenesis and therapeutic strategies.
Purpose of the Study:
- To investigate the therapeutic effects of Fasudil, a Rho kinase (ROCK) inhibitor, in a mouse model of EAE.
- To elucidate the underlying mechanisms of Fasudil's action, focusing on immune cell modulation and macrophage polarization.
Main Methods:
- Fasudil was administered intraperitoneally to EAE mice during early and late disease stages.
- ROCK-II expression, T cell subsets (CD4+IL-17+, CD4+IL-10+), and macrophage polarization markers (M1/M2) in spleen and spinal cord were analyzed.
- Western blot and immunohistochemistry were employed to assess specific protein expression (iNOS, Arg-1).
- In vitro experiments evaluated Fasudil's direct effect on macrophage phenotype.
Main Results:
- Fasudil significantly ameliorated clinical symptoms of EAE.
- Treatment reduced ROCK-II expression and inflammatory cell infiltration, improving demyelination.
- Fasudil modulated T cell responses, inhibiting CD4+IL-17+ cells early and promoting CD4+IL-10+ regulatory T cells later.
- A key finding was the shift from M1 to M2 macrophages, evidenced by altered expression of M1 (CD16/32, iNOS, IL-12, TLR4, CD40) and M2 (CD206, Arg-1, IL-10, CD14) markers.
- In vitro studies confirmed Fasudil's ability to induce M2 polarization independently of other cells.
- M2 polarization correlated with reduced pro-inflammatory cytokines (IL-1β, TNF-α, MCP-1).
Conclusions:
- Fasudil demonstrates significant therapeutic potential for EAE.
- The mechanism involves suppressing inflammatory responses and promoting M2 macrophage polarization.
- Fasudil's immunomodulatory effects suggest it could be a valuable therapeutic agent for MS and related inflammatory neurological conditions.
