Xanthium strumarium L. extract attenuates TNF-α/IFN-γ- stimulated inflammation and promotes hyaluronan production on HaCaT keratinocytes.

BACKGROUND: Xanthium strumarium L. has traditionally been used for inflammatory disorders; however, its molecular effects on keratinocyte inflammation and hyaluronan metabolism remain poorly understood. This study evaluated the modulatory trends of X.strumarium L. extract (XsE) on TNF-α/IFN-γ-stimulated inflammatory signaling pathway and hyaluronan regulation on HaCaT keratinocytes. METHODS AND RESULTS: The antioxidative stress of XsE was evaluated by free radical scavenging assays and dichlorofluorescein fluorescence staining. XsE exhibited significant antioxidant activity without inducing cytotoxicity in HaCaT cells and markedly attenuated TNF-α/IFN-γ-induced ROS accumulation. The gene expression and ELISA assays revealed that XsE suppressed pro-inflammatory cytokines and chemokines, including IL-6, IL-8, TARC, RANTES, and CTACK, at both transcriptional and protein levels. Mechanistically, Western blot analysis demonstrated that XsE treatment was strongly associated with the suppression of p38, IκBα, and p65 phosphorylation within the canonical NF-κB and specific p38 MAPK signaling cascades. In addition, XsE enhanced hyaluronan accumulation by upregulating hyaluronan synthase gene transcription, resulting in verified downstream macromolecule secretion. UPLC-MS analysis identified chlorogenic acid, miquelianin, hyperoside, cynarine, and syringetin as major phytochemical constituents in XsE. Molecular docking analysis further supported the predicted interactions of these compounds with inflammation- and hyaluronan-related targets. CONCLUSIONS: These findings demonstrate that the anti-inflammatory and hyaluronan-producing properties of XsE may contribute to the regulation of epidermal inflammatory responses and skin barrier-associated functions.