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A high\u2010fat diet (HFD) is a key contributing factor, closely associated with gut microbiota dysbiosis. Although immunomodulatory effects of\n                      <jats:italic>Ganoderma lucidum<\/jats:italic>\n                      spore oil (GLSO) have been reported, its role in lipid metabolism disorders and its interaction with gut microbiota remain incompletely understood.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>RESULTS<\/jats:title>\n                    <jats:p>\n                      In this study, an HFD\u2010induced obese mouse model was established and treated with GLSO (800\u2009mg\u2009kg\n                      <jats:sup>\u22121<\/jats:sup>\n                      \u2009day\n                      <jats:sup>\u22121<\/jats:sup>\n                      ) for 82\u2009days. Integrated analyses \u2013 including 16S rRNA sequencing, metabolomics, fecal fermentation of GLSO (FGLSO), and HepG2 cell experiments \u2013 were performed to evaluate metabolic effects and potential microbiota\u2010related mechanisms.\n                      <jats:italic>Ganoderma lucidum<\/jats:italic>\n                      spore oil significantly reduced body weight gain, fasting blood glucose, serum lipid levels \u2013 including total cholesterol (TC), triglycerides (TG), and low\u2010density lipoprotein cholesterol (LDL\u2010C) \u2013 and inflammatory markers, specifically tumor necrosis factor\u2010alpha (TNF\u2010\n                      <jats:italic>\u03b1<\/jats:italic>\n                      ), interleukin\u20101 beta (IL\u20101\n                      <jats:italic>\u03b2<\/jats:italic>\n                      ) and interleukin\u20106 (IL\u20106). It also improved gut microbiota composition by increasing microbial diversity and enriching beneficial taxa such as\n                      <jats:italic>Bacillus<\/jats:italic>\n                      and unclassified_Muribaculaceae. These changes were accompanied by increased levels of short\u2010chain fatty acids (SCFAs), particularly acetate and propionate, as well as alterations in bile acid metabolism. Further analysis suggested that metabolites may be associated with the regulation of hepatic lipid metabolism, potentially involving the peroxisome proliferator\u2010activated receptor alpha (PPAR\n                      <jats:italic>\u03b1<\/jats:italic>\n                      ) signaling pathway. Fecal fermentation of GLSO exhibited stronger lipid\u2010lowering effects than native GLSO\n                      <jats:italic>in vitro<\/jats:italic>\n                      , indicating that gut microbiota\u2010mediated biotransformation may enhance the bioactivity of GLSO.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>CONCLUSION<\/jats:title>\n                    <jats:p>Collectively, these findings suggest that GLSO alleviates HFD\u2010induced metabolic disorders and highlight a potential role of gut microbiota in modulating its metabolic effects and enhancing its bioactivity. \u00a9 2026 Society of Chemical Industry.<\/jats:p>\n                  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