3‐Oxo‐22α‐Hydroxy‐Rotundic Acid Alleviates Hyperlipidemia in Mice by Modulating Lipid Metabolism Through the <scp>AMPK</scp>‐<scp>SREBP</scp>‐1c‐<scp>PPARα</scp> Pathway

3-Oxo-22α-hydroxy-rotundic acid (ITP3) demonstrated notable hypolipidemic activity. However, the molecular mechanism of its hypolipidemic activity has not been elucidated. The present study aimed to evaluate its lipid-lowering efficacy using in vivo and in vitro hyperlipidemia models and to further elucidate its potential mechanism of action in hyperlipidemia. Endophytic fungi in plants of the genus Ilex were utilized for microbial transformation of rotundic acid (RA) to generate an adequate quantity of ITP3. Free fatty acid (FFA) treatment of HepG2 cells and C57BL/6J mice was used to evaluate the hypolipidemic effects of ITP3 in vivo and in vitro. A metabolomics approach combined with Western blot analysis was used to reveal the potential mechanism of the anti-hyperlipidemia of ITP3. The results showed that ITP3 exhibited good lipid-lowering activity in vivo and in vitro models of hyperlipidemia. In addition, metabolomics analysis revealed significant changes in serum and intracellular metabolite lipid levels, which were restored by ITP3. Mechanistically, ITP3 can inhibit lipid synthesis and activate lipid oxidation via the AMPK-SREBP-1c-PPARα pathway, thereby ameliorating lipid metabolism disorders. ITP3 exhibits a promising lipid-lowering effect via the AMPK-SREBP-1c-PPARα pathway, thereby improving lipid metabolism. This work highlights ITP3 as a potential phytochemical candidate for the treatment of hyperlipidemia.