Atractylenolide III alleviates MASLD by inhibiting lipid synthesis and apoptosis via PTEN.
Abstract
Background
Metabolic dysfunction-associated steatotic liver disease (MASLD) has emerged as a major global health issue. This study aims to investigate the ameliorative effects of atractylenolide III (ATL III) on MASLD and its underlying molecular mechanisms, thereby providing theoretical basis and experimental support for the development of therapeutic drugs for MASLD.
Methods
A high-fat diet (HFD)-fed mouse model and a combination of oleic acid and palmitic acid (PO)-induced lipid deposition model in AML-12 hepatocytes were established. The therapeutic effects of ATL III on hepatic lipid accumulation, injury, and inflammation were evaluated using biochemical assays, histopathological staining, immunofluorescence, Western blot (WB), and quantitative real-time PCR. Transcriptome sequencing was performed to screen for key targets, and molecular docking together with cellular thermal shift assay (CETSA) were used to validate the potential binding between ATL III and the target protein. The findings were further verified using a liver-specific Pten knockdown mouse model.
Results
In vivo experiments showed that ATL III significantly ameliorated hepatic lipid accumulation, injury, and inflammatory response in HFD feeding mice. In vitro experiments demonstrated that ATL III effectively reduced lipid accumulation in PO-induced AML-12 cells without affecting cell viability. Transcriptome sequencing and KEGG enrichment analysis revealed that the differentially expressed genes regulated by ATL III were enriched in the phosphatidylinositol 3‑kinase (PI3K)/serine/threonine protein kinase B (AKT), fatty acid metabolism, and apoptosis pathways. Further studies showed that ATL III engaged PTEN in a cellular context and upregulated its expression, thereby inhibiting excessive AKT activation. WB analysis confirmed that ATL III regulated lipid synthesis and apoptosis-related proteins via the PTEN/AKT axis. Notably, liver‑specific knockdown of Pten significantly attenuated the ameliorative effects of ATL III on the expression of the above molecules, as well as on liver injury and lipid accumulation.
Conclusions
ATL III ameliorates hepatic steatosis, injury, and inflammation in HFD-induced MASLD mice. Mechanistically, ATL III upregulates PTEN expression and suppresses excessive AKT activation, thereby reducing hepatic lipid synthesis and hepatocyte apoptosis. Liver-specific knockdown of Pten attenuates the protective effects of ATL III, confirming that PTEN is functionally required for its beneficial actions. These findings suggest that ATL III represents a potential therapeutic candidate for MASLD by acting through the PTEN/AKT pathway.