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MASLD is a multifactorial condition, which still needs to be completely understood. Galectin 3 (Gal-3) is up-regulated in several liver disorders suggesting its implication in the mechanisms underlying liver damage.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec026\">\n<jats:title>Methods<\/jats:title>\n<jats:p>A human multilineage 3D model was utilized to investigate the role of Gal-3 in MASLD development. Human hepatoma cell line (HepG2) and human stellate cell line (LX-2) were co-cultured in a physiological ratio of 24:1 and treated with a mixture of palmitic and oleic acid (PAOA, ratio 1:2) to induce hepatocyte steatosis and facilitate the development of fibrosis. While the effect of <jats:italic>LGALS3<\/jats:italic> silencing on neutral fat content was assessed by Oil-Red-O (ORO) staining, type I collagen production was analysed by immunofluorescent staining for collagen type I alpha 1 (COL1A1).<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec027\">\n<jats:title>Results<\/jats:title>\n<jats:p>Gal-3 depletion caused a reduction of neutral lipid content and COL1A1 accumulation in 3D spheroids. While <jats:italic>LGALS3<\/jats:italic> silencing did not significantly alter the respiratory state, analysis of genes involved in lipid metabolism demonstrated significant changes in genes involved in \u03b2-oxidation and triglyceride synthesis.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec028\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>These results suggest a role of Gal-3 in the regulation of fatty acid and collagen accumulation, thereby indicating that approaches aimed at inhibiting Gal-3 may represent a promising therapeutic strategy in MASLD.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0326373","type":"journal-article","created":{"date-parts":[[2025,7,3]],"date-time":"2025-07-03T17:21:59Z","timestamp":1751563319000},"page":"e0326373","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Galectin-3 inhibition ameliorates hepatic steatosis in a multilineage 3D spheroid 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