Background: Dihydroartemisinin (DHA), a bioactive sesquiterpenoids originated from Artemisia annua L. , extensively applied in traditional medicine for the treatment of multiple autoimmune diseases. AXL, shown to dampen the immune response, reduce pro-inflammatory cytokine secretion, clear apoptotic cells and debris in Multiple sclerosis (MS), suggesting that it plays a role in disease pathogenesis. Thus, aiming to investigated whether DHA ameliorates experimental autoimmune encephalomyelitis (EAE) by modulating AXL signaling, we determined it both in vivo and vitro studies.Methods: Firstly, female C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein MOG 35-55 peptide emulsified in complete Freund’s adjuvant and injected with pertussis toxin on day 0 and 2. Mice were monitored daily with DHA for clinical signs of disease and analyzed for pathology and ethology during the acute phase of disease. Moreover, functions of Immunological responses were assessed by flow cytometry and fluorescence microscope, chemokine were determined through ELISA and Transwell assay, proteins were monitored by Western blot and genes expression were explored by PCR.Results: Results identified mice immunized with MOG35-55 had a significantly more severe acute phase of EAE than negative control mice. In contrast, DHA treated mice had slighter spinal cord lesions with less inflammatory cuffs, less demyelination, and slighter axonal damage than EAE mice. Furthermore, by using gait analysis, DHA obviously ameliorated clinical features in EAE mice. Strikingly, DHA reduced the chemotactic ability of microglia, promoted the differentiation of T cells into Treg cells and enhanced the phagocytic ability of microglia. Moreover, DHA up-regulated expression of AXL, SOCS3, and STAT1 phosphorylation. By contrast, no significant differences were noted in these responses above by using SGI7079, inhibitor of AXL. Furthermore, DHA up-regulated expression of AXL, SOCS3, and STAT1 phosphorylation and enhanced the phagocytic ability of microglia when IFNAR was activated by IFN-β,however, no significant differences were noted in these responses above by using Fludarabine, inhibitor of STAT1 phosphorylation.Conclusions: Taken together, the present study demonstrated that DHA exerts multiple effects in ameliorating neuroinflammation process by enhancing AXL signaling that depending on modulating STAT1/SOCS3 axis in EAE, a mouse model of multiple sclerosis.