<p>Demyelination in multiple sclerosis (MS) is associated with chronic inflammation and dysregulation of cholesterol metabolism. In this study, we investigated the protective effects of silymarin, an herbal flavonoid, against inflammation, impairment of brain cholesterol metabolism, and demyelination in the experimental autoimmune encephalomyelitis (EAE) model of MS. Given the well-documented anti-hypercholesterolemic properties of silymarin, we sought to assess its protective effects in combination with a cholesterol-supplemented diet. Additionally, the effects of the cholesterol-supplemented diet alone were evaluated. Female C57BL/6 mice were fed either a standard chow diet or a cholesterol-supplemented standard chow for 2&#xa0;weeks prior to EAE induction, continuing until day 20 post-immunization. Experimental groups received silymarin, a cholesterol-supplemented diet, or both treatments. Silymarin significantly reduced clinical symptoms and the incidence of EAE, alleviated inflammation, prevented severe demyelination, modulated the expression genes involved in brain cholesterol metabolism, and ultimately improved the serum lipid profile. The cholesterol-supplemented diet alone was partially effective. Co-administration of silymarin with the cholesterol-supplemented diet showed superior efficacy across some outcomes. String web analysis revealed the significant interaction of apolipoprotein E (ApoE), a major component of brain HDL-like particles, with other key genes. ELISA results demonstrated that silymarin alone and in combination with the cholesterol-supplemented diet restored ApoE levels in EAE-induced mice. These findings suggest that silymarin, in conjunction with a cholesterol-supplemented diet, warrants further investigation in clinical trials as a potential protective strategy for MS management.</p>