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<i>Gentiana lutea</i> is a bitter herb that is traditionally used to improve gastric disorders. Recently, we have shown that <i>Gentiana lutea</i> extract (GE) also modulates the lipid metabolism of human keratinocytes in vitro and in vivo. In the present study, we investigated the role of GE on ceramide synthesis in human primary keratinocytes (HPKs) and psoriasis-like keratinocytes. We could demonstrate that GE increased the concentrations of glucosylceramides and the ceramide AS/AdS subclass without affecting the overall ceramide content in HPKs. The expression of ceramide synthase 3 (<i>CERS3</i>) and elongases (<i>ELOVL1</i> and <i>4</i>) was reduced in psoriasis lesions compared to healthy skin. Psoriasis-like HPKs, generated by stimulating HPKs with cytokines that are involved in the pathogenesis of psoriasis (IL-17, TNF-α, IL-22 and IFN-γ) showed increased levels of IL-6, IL-8 and increased expression of <i>DEFB4A</i>, as well as decreased expression of <i>ELOVL4</i>. The treatment with GE partly rescued the reduced expression of <i>ELOVL4</i> in psoriasis-like HPKs and augmented <i>CERS3</i> expression. This study has shown that GE modulates ceramide synthesis in keratinocytes. Therefore, GE might be a novel topical treatment for skin diseases with an altered lipid composition such as psoriasis.