Xanthoangelol-enriched Angelica keiskei Koidzumi targets foamy Kupffer cells to ameliorate MASLD by activating the TFEB-mediated CLEAR network

Background Metabolic dysfunction-associated steatotic liver disease (MASLD) progression is driven by lipid overload and inflammatory activation of Kupffer cells (KCs). Xanthoangelol (XAG), a major chalcone in Angelica keiskei Koidzumi, has anti-inflammatory activity, but its effects on KC immunometabolic dysfunction remain unclear. Objective To determine whether XAG-enriched A. keiskei extract ameliorates MASLD by targeting foamy KC dysfunction. Methods MASLD was induced in male C57BL/6 N mice by fast-food diet feeding for 15 weeks, followed by daily oral administration of XAG-enriched A. keiskei extract for a further 9 weeks. Liver injury, steatosis, collagen deposition, inflammatory signaling, and KC abundance were evaluated. Primary hepatocytes, primary KCs, immortalized KCs, and reporter macrophages were used to define cell-specific responses, and KC-to-hepatocyte crosstalk was assessed by conditioned-medium transfer. MASH-like KC dysfunction was modeled using palmitic acid, oleic acid, LPS, glucose, fructose, and TGF-β with or without XAG. RNA sequencing, pathway enrichment, immunoblotting, RT-qPCR, and immunofluorescence were used to assess inflammatory, lipid-handling, autophagy–lysosome, and TFEB–CLEAR pathways. Results XAG-enriched A. keiskei extract reduced FFD-induced liver injury, steatosis, collagen deposition, metabolic dysfunction, inflammatory signaling, and KC accumulation. Primary hepatocytes showed limited responses in the lipid-related endpoints examined, whereas KCs exhibited pronounced inflammatory and immunometabolic responses to XAG. In MASH-stimulated KCs, XAG suppressed NF-κB/MAPK-associated inflammatory activation, reduced lipid accumulation, attenuated NLRP3 inflammasome signaling, and activated autophagy–lysosome and lipid-clearance programs. Mechanistically, XAG promoted TFEB nuclear localization and increased CLEAR network gene expression. Conclusion XAG-enriched A. keiskei extract ameliorates experimental MASLD by suppressing foamy KC inflammatory activation and enhancing TFEB–CLEAR-associated lysosomal regulation, supporting XAG as a KC-directed immunomodulatory candidate.