Wedelolactone targets SORBS1 to regulate glycolipid metabolism and suppress inflammation, thereby alleviating diet-induced MASH via multiple pathways.

BACKGROUND

The pathogenic mechanisms underlying metabolic dysfunction-associated steatohepatitis (MASH) are highly complex and multifactorial, and there are limited pharmacological therapies proven effective for clinical use. Although the natural product wedelolactone (WED) demonstrates potential hepatoprotective effects, its therapeutic efficacy in MASH and the underlying mechanisms remain poorly understood.

PURPOSE: This study investigates the therapeutic efficacy of WED in ameliorating diet-induced MASH and aims to elucidate its direct molecular targets and underlying mechanisms.

METHODS

To investigate the therapeutic potential of WED in MASH, this study established an in vitro model of lipid accumulation and inflammatory response by treating HepG2 cells and primary mouse hepatocytes with palmitic acid (PA) and oleic acid (OA), thereby assessing the in vitro effectiveness of WED. Additionally, we employed the MCD diet- and HFHC diet-induced mouse MASH models to evaluate the therapeutic effects of WED. To identify its molecular targets, we synthesized biotin-labeled WED probes and screened potential direct binding proteins using a human proteome microarray. The binding affinity was validated by surface plasmon resonance (SPR) and cellular thermal shift assay (CETSA). Molecular dynamics simulation and molecular docking were performed to determine the key interaction sites between WED and its targets. Transcriptomic and bioinformatics analyses were integrated to elucidate the downstream signaling pathways regulated by WED through these targets. Finally, biological functional assays were performed to validate the underlying mechanism of WED.

RESULTS

This study has demonstrated that WED exerts a significant anti-MASH effect by directly targeting SORBS1, with binding occurring at VAL32 and HIS47. The underlying mechanism involves the direct binding and activation of SORBS1 by WED, leading to multiple regulation of downstream signaling pathways. Specifically, WED regulates the expression of SORBS1 in liver tissue, improves glycolipid metabolism through the CBL/TC10/GLUT4 axis, and suppresses proinflammatory cytokines such as TNF-α and IL-6. This coordinated action results in effective alleviation of hepatic lipid accumulation, inflammation, and fibrosis. Furthermore, inhibiting SORBS1 exacerbates the MASH process and completely abolishes the efficacy of WED, thereby confirming that SORBS1 is the key target for its therapeutic effect.

CONCLUSION

Here, this study reveals that WED ameliorates insulin resistance and suppresses hepatic inflammation and fibrosis by directly binding to SORBS1, thereby significantly attenuating the progression of MASH. These findings establish SORBS1 as a potential novel therapeutic target for MASH and provide a potential candidate compound for the advancement of therapies targeting MASH.