Multiomics Analysis Reveals Stevioside Alleviates MASLD via Modulating Gut Microbiota and Suppressing AGEs‐RAGE/NLRP3 Axis
Feng Zhang, Xueyun Dong, Hao Xu, Xuehui Liu, Yunhan Xie, Ruijia Qian, Yingting Qian, Liyun Ling, Minchao Ye, Asmaa Ali, Liang Wu, Keke ShaoBackground
Effective pharmacotherapies for metabolic dysfunction–associated steatotic liver disease (MASLD), a condition tightly linked to metabolic dysregulation, remain scarce. We aimed to elucidate how stevioside (ST) ameliorates MASLD via the gut microbiota–liver axis and the advanced glycation endproducts (AGEs)‐RAGE/NLRP3 pathway.
Methods
C57BL/6 mice were fed a high‐fat diet (HFD) for 12 weeks to establish MASLD and then treated with low‐ (50 mg/kg) or high‐dose (200 mg/kg) ST. We employed histological staining, biochemical assays (ELISA, qPCR, and Western blot), 16 S rRNA gene sequencing, metabolomics, and network pharmacology to analyze the effects.
Results
ST treatment significantly reversed HFD‐induced weight gain, steatosis, and hepatic inflammation. It reduced proinflammatory cytokines and suppressed the AGEs‐RAGE/NLRP3 axis. At the molecular level, ST enhanced PPARα expression while decreasing the expression of SREBP‐1c and NLRP3. Gut microbiota analysis showed an enrichment of butyrate‐producing Bacteroides_H and Ligilactobacillus , alongside a reduction in Lawsonibacter . This shift increased fecal butyrate, upregulated gut barrier proteins (ZO‐1 and Occludin), and prevented LPS leakage. Metabolomics highlighted changes in lipid mediators (increased 5‐OxoETE/EpDPA and decreased mevalonate), suggesting optimized cholesterol and arachidonic acid metabolism. Network pharmacology linked ST targets (JUN and STAT3) to key metabolic pathways.
Conclusion
ST mitigates MASLD by synergistically modulating the “microbiota–SCFAs–gut barrier” integrity and suppressing AGEs‐RAGE/NLRP3 signaling. This provides a theoretical rationale for utilizing ST in functional foods or clinical therapies for MASLD.