Oligodendrocyte (OL) death and remyelination failure lead to progressive neurological deficits in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Matrine (MAT), a quinolizidine alkaloid component derived from the root of Sophora flavescens, has the capacity to effectively inhibit central nervous system (CNS) inflammation and to promote neuroregeneration. In the present study we explored its regulatory mechanism on the Wnt/beta-catenin/TCF7L2 pathway, a negative modulator for myelination, in MOG35-55 peptide-induced EAE. Our results clearly indicate that MAT treatment reduced the activation of Wnt3a and beta-catenin in the CNS of EAE mice, accompanied by the activation of GSK3 beta and decreased expression of cyclin D1 and Axin2, two target genes of the Wnt3a/beta-catenin pathway. In addition, MAT increased OL maturation and myelination, as evidenced by the decreased number of NG(2+)Olig(2+) cells and the increased numbers of MBP+ and CC(1+)Olig(2+) cells. Taken together, these findings indicate that MAT treatment promoted the maturation of OLs and myelin repair, which is closely related to the modulation of the Wnt/beta-catenin/TCF7L2 signaling pathway.