Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system characterized by recurrent and progressive demyelination/remyelination cycles, neuroinflammation, oligodendrocyte loss, and axonal pathology. Baicalein isolated from the roots of Scutellaria baicalensis has been shown to exert anti-inflammatory and antioxidant effects. The cuprizone model is an established mouse model of MS and causes demyelination and motor dysfunction and induces neuroinflammation, such as glial activation and pro-inflammatory cytokine production. To determine whether Baicalein attenuates cuprizone-induced demyelination, we administrated Baicalein to cuprizone-exposed mice. Baicalein attenuated weight loss (P < 0.05) and motor dysfunction (P < 0.05) in the cuprizone model mice. Baicalein treatment effectively suppressed the demyelination (P < 0.01) and gene expressions of CNP (P < 0.05) and MBP (P < 0.05). Baicalein treatment also inhibited the cuprizone-induced increase in Ibal-positive microglia (P < 0.001), GFAP-positive astrocytes (P < 0.001), and the gene expressions of CD11b (P < 0.01), GFAP (P < 0.05), TNFa (P < 0.05), IL-113 (P < 0.05), and iNOS (p < 0.01). We found that Baicalein treatment attenuated cuprizone-induced demyelination, glial activation, pro-inflammatory cytokine expression, and motor dysfunction. Our results suggest that Baicalein may be a useful therapeutic agent in demyelinating diseases to suppress neuroinflammation.