Fasudil regulates T cell responses through polarization of BV-2 cells in mice experimental autoimmune encephalomyelitis  

作　　者：Chan CHEN Yan-hua LI Qiong ZHANG Jie-zhong YU Yong-fei ZHAO Cun-gen MA Bao-guo XIAO 

机构地区：[1]Institute of Neurology, Huashan Hospital, Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai 200040, China [2]Institute of Brain Science, Department of Neurology, Medical School, Shanxi Datong University, Datong 037009, China [3]Department of Encepha/opathy and National Major C/inica/ Department of Ministry of Health, Third Hospital, Shanxi University of Traditiona/ Chinese Medicine, Taiyuan 030006, China

出　　处：《Acta Pharmacologica Sinica》2014年第11期1428-1438,共11页中国药理学报（英文版）

基　　金：Acknowledgements This work was supported by grants from the National Natural Science Foundation of China （No 81070957, 81371414, and 81070956）.

摘　　要：Aim： Fasudil, a selective Rho kinase （ROCK） inhibitor, has been shown to alleviate the severity of experimental autoimmune encephalomyelitis （EAE） via attenuating demyelination and neuroinflammation. The aim of this study was to investigate the effects of fasudil on interactions between macrophages/microglia and T cells in a mice EAE model. Methods： Mouse BV-2 microglia were treated with IFN-γ and fasudil. Cell viability was detected with MTT assay. BV-2 microglia polarization was analyzed using flow cytometry. Cytokines and other proteins were detected with ELISA and Western blotting, respectively. Mice were immunized with MOG35-55 to induce EAE, and then treated with fasudil （40 mg/kg, ip） every other day from d 3 to d 27 pi. Encephalomyelitic T cells were prepared from the spleen of mice immunized with MOG35-55 on d 9 pi. Results： Treatment of mouse BV-2 microglia with fasudil （15 μg/mL） induced significant phenotype polarization and functional plasticity, shifting M1 to M2 polarization. When co-cultured with the encephalomyelitic T cells, fasudil-treated BV-2 microglia significantly inhibited the proliferation of antigen-reactive T cells, and down-regulated IL-17-expressing CD4＋ T cells and ILo17 production. Furthermore, fasudil-treated BV-2 microglia significantly up-regulated CD4＋CD25high and CD4＋IL-10＋ regulatory T cells （Tregs） and IL-IO production, suggesting that the encephalomyelitic T cells had converted to Tregs. In EAE mice, fasudil administration significantly decreased both CD11b＋iNOS＋ and CD11b＋NF-α＋ M1 microglia, and increased CD11b＋IL-10＋ M2 microglia. Conclusion： Fasudil polarizes BV-2 microglia into M2 cells, which convert the encephalomyelitic T cells into Tregs in the mice EAE model.Aim： Fasudil, a selective Rho kinase （ROCK） inhibitor, has been shown to alleviate the severity of experimental autoimmune encephalomyelitis （EAE） via attenuating demyelination and neuroinflammation. The aim of this study was to investigate the effects of fasudil on interactions between macrophages/microglia and T cells in a mice EAE model. Methods： Mouse BV-2 microglia were treated with IFN-γ and fasudil. Cell viability was detected with MTT assay. BV-2 microglia polarization was analyzed using flow cytometry. Cytokines and other proteins were detected with ELISA and Western blotting, respectively. Mice were immunized with MOG35-55 to induce EAE, and then treated with fasudil （40 mg/kg, ip） every other day from d 3 to d 27 pi. Encephalomyelitic T cells were prepared from the spleen of mice immunized with MOG35-55 on d 9 pi. Results： Treatment of mouse BV-2 microglia with fasudil （15 μg/mL） induced significant phenotype polarization and functional plasticity, shifting M1 to M2 polarization. When co-cultured with the encephalomyelitic T cells, fasudil-treated BV-2 microglia significantly inhibited the proliferation of antigen-reactive T cells, and down-regulated IL-17-expressing CD4＋ T cells and ILo17 production. Furthermore, fasudil-treated BV-2 microglia significantly up-regulated CD4＋CD25high and CD4＋IL-10＋ regulatory T cells （Tregs） and IL-IO production, suggesting that the encephalomyelitic T cells had converted to Tregs. In EAE mice, fasudil administration significantly decreased both CD11b＋iNOS＋ and CD11b＋NF-α＋ M1 microglia, and increased CD11b＋IL-10＋ M2 microglia. Conclusion： Fasudil polarizes BV-2 microglia into M2 cells, which convert the encephalomyelitic T cells into Tregs in the mice EAE model.

关 键 词：FASUDIL multiple sclerosis experimental autoimmune encephalomyelitis NEUROINFLAMMATION MICROGLIA T cell cellpolarization CYTOKINE 

分 类 号：Q2[生物学—细胞生物学] TN16[电子电信—物理电子学]