Targeting the shift from M1 to M2 macrophages in experimental autoimmune encephalomyelitis mice treated with fasudil

• Published in: PloS one Vol. 8; no. 2; p. e54841
• Main Authors: Liu, Chunyun, Li, Yanhua, Yu, Jiezhong, Feng, Ling, Hou, Shaowei, Liu, Yueting, Guo, Mingfang, Xie, Yong, Meng, Jian, Zhang, Haifei, Xiao, Baoguo, Ma, Cungen
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 13.02.2013; Public Library of Science (PLoS)
• Subjects: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - therapeutic use; Animal experimentation; Animals; Antigens; Biology; CD14 antigen; CD16 antigen; CD4 antigen; CD40 antigen; Cell adhesion & migration; Chinese medicine; Cords; Demyelination; Encephalomyelitis; Encephalomyelitis, Autoimmune, Experimental - drug therapy; Encephalomyelitis, Autoimmune, Experimental - immunology; Encephalomyelitis, Autoimmune, Experimental - metabolism; Enzyme inhibitors; Experimental allergic encephalomyelitis; Female; Health aspects; Immunohistochemistry; Immunoregulation; Inflammation; Inhibition; Interleukin 10; Interleukin 12; Interleukin 17; Interleukin-10 - metabolism; Laboratory animals; Lymphocytes; Lymphocytes T; Macrophages; Macrophages - drug effects; Macrophages - immunology; Macrophages - metabolism; Medical schools; Medicine; Mice; Monocyte chemoattractant protein 1; Multiple sclerosis; Neurology; Neurosciences; Nitric Oxide Synthase Type II - metabolism; Nitric-oxide synthase; Pathogenesis; Polarization; Protein Kinase Inhibitors - pharmacology; Protein Kinase Inhibitors - therapeutic use; Rho-associated kinase; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - metabolism; Rodents; Science; Severity of Illness Index; Spleen; Spleen - drug effects; Spleen - immunology; Spleen - metabolism; T cell receptors; T cells; T-Lymphocytes - drug effects; T-Lymphocytes - metabolism; TLR4 protein; Toll-like receptors; Tumor necrosis factor; Tumor Necrosis Factor-alpha - metabolism; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0054841

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.