title: [Immunoregulative effect of Fasudil on encephalomyelitic T cells in experimental autoimmune encephalomyelitis mice].
abstract: OBJECTIVE
  To explore the therapeutic effect of Fasudil-modified splenic mononuclear cells (MNCs) in experimental autoimmune encephalomyelitis (EAE) and the possible mechanisms.
  
  
  METHODS
  C57BL/6 female mice were immunized with myelin oligodendrocyte glycoprotein peptide 35-55 to establish active immunity EAE model. Splenic MNCs were isolated on the 9th day after immunization and treated with or without Fasudil for 72 h in vitro. These cells were collected for analysis of the variance of T cell subtypes, the level of cytokines and the activity of Rho kinase (ROCK). MNCs (5×107 cells) were resuspended in 500 µL of phosphate buffer solution (PBS) and transferred into EAE model (intraperitoneal injection), which was divided into a PBS-MNCs group and a Fasudil-MNCs group. Changes of body weight and clinical symptom scores were observed.
  
  
  RESULTS
  Splenic encephalitogenic MNCs from EAE mice on the 9th day after immunization could establish passive transfer EAE model. But Fasudil-treated MNCs did not trigger EAE development. Compared with the PBS-MNCs group, the loss of body weight was less in the Fasudil-MNCs group. The in vitro experiment indicated that Fasudil could suppress the activity of ROCK on MNCs (P<0.01), decrease the percentage of CD4+ T cells with the expression of interferon-γ (IFN-γ) and interleukin-17 (IL-17) (IFN-γ: P<0.01; IL-17: P<0.05), while increase the secretion of CD4+ T cells with the expression of transforming growth factor-β (TGF-β) and IL-10 (all P<0.001) . Furthermore, Fasudil could inhibit the release of IL-17 (P<0.001) and enhance the level of IL-10 (P<0.05).
  
  
  CONCLUSIONS
  Fasudil-modified cell therapy affects the occurrence and development of EAE by inhibiting the inflammatory reaction of helper T cell 1 (Th1) and Th17 while enhancing the immunoregulative effect of Th2.
authors.affiliation: Institute of Brain Science, Shanxi Datong University, Datong Shanxi 037009, China.
authors.forename: Chunyun
authors.initials: C
authors.lastname: Liu
authors.affiliation: Institute of Brain Science, Shanxi Datong University, Datong Shanxi 037009, China.
authors.forename: Shangde
authors.initials: S
authors.lastname: Guo
authors.affiliation: Institute of Brain Science, Shanxi Datong University, Datong Shanxi 037009, China.
authors.forename: Nianping
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Institute of Brain Science, Shanxi Datong University, Datong Shanxi 037009, China.
authors.forename: Jiezhong
authors.initials: J
authors.lastname: Yu
authors.affiliation: Institute of Neurology, Huashan Hospital, Fudan University, Shanghai 200040, China.
authors.forename: Baoguo
authors.initials: B
authors.lastname: Xiao
authors.affiliation: Institute of Brain Science, Shanxi Datong University, Datong Shanxi 037009;"2011" Collaborative Innovation Center, Shanxi University of Traditional Chinese Medicine, Taiyuan 030024, China.
authors.forename: Cungen
authors.initials: C
authors.lastname: Ma
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives; D000818:Animals; D004681:Encephalomyelitis, Autoimmune, Experimental; D005260:Female; D007371:Interferon-gamma; D016753:Interleukin-10; D020381:Interleukin-17; D051379:Mice; D008810:Mice, Inbred C57BL; D063308:Myelin-Oligodendrocyte Glycoprotein; D013154:Spleen; D013601:T-Lymphocytes; D016212:Transforming Growth Factor beta; D054460:rho-Associated Kinases
chemical_list: C556062:IL10 protein, mouse; D020381:Interleukin-17; D063308:Myelin-Oligodendrocyte Glycoprotein; D016212:Transforming Growth Factor beta; D016753:Interleukin-10; D007371:Interferon-gamma; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D054460:rho-Associated Kinases; C049347:fasudil