Hydroxyfasudil ameliorates experimental autoimmune encephalomyelitis possibly through immunomodulation and anti-inflammatory effects

Abstract

Background Rho kinase inhibitor Fasudil is effective in the treatment of EAE, but its narrow safety window, lower bioavailability limit the clinical use. Hydroxyfasudil, active metabolite of Fasudil in vivo, exhibits higher biological activity and longer half-period. Objective To observe the therapeutic potential of Hydroxyfasudil in EAE, and explore its possible mechanisms. Material and methods The effects of Hydroxyfasudil on cell viability. We further explored therapeutic effect in EAE model. Female C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein peptides (MOG35-55), which were divided into EAE group, Hydroxyfasudil group. Hydroxyfasudil was injected intraperitoneally (40 mg/kg/day) on day 3 post immunization. The injection of saline was set up as control in a similar manner. Results Hydroxyfasudil was well tolerant in vitro. Hydroxyfasudil delayed the clinical onset and suppressed the severity of EAE. Demyelination and cell infiltration in spinal cords of Hydroxyfasudil group were significantly decreased. Flow cytometry analysis showed that Hydroxyfasudil reduced CD4+IFN-γ+ and CD4+IL-17+ but increased CD4+CD25+ T cells. Hydroxyfasudil decreased inflammatory cytokine, chemokine and inhibited NF-κB, COX-2, iNOS. According to RT-PCR analysis, Hydroxyfasudil suppressed iNOS, CD68 and TNF-α, while enhanced Arg-1, IL-4 and TGF-β. Conclusion Hydroxyfasudil should be a potential approach for EAE through immunomodulation, anti-inflammatory effects, the transformation of M1 type macrophages to M2, preventing the infiltration of inflammatory cells into CNS. (NNSF of China 81473577, Shanxi Scholarship Council of China 2014-7, 2011 Cultivation Project of Shanxi University of TCM 2011PY-1. Ma and Xiao are corresponding authors).