An Integrated Multi‐Omics Analysis Reveals the Protective Mechanism of Aspirin on Metabolic Dysfunction‐Associated Steatotic Liver Disease

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a growing health concern worldwide. Aspirin has shown potential in ameliorating MASLD, yet its mechanisms remain incompletely understood. This study aims to investigate the protective effects of aspirin on MASLD by incorporating transcriptomic and metabolomic approaches. Mice were fed a high-fat diet (HFD) to induce MASLD and treated with aspirin for 12 weeks. Blood and liver samples were collected for biochemical assays, histological analysis, RT-qPCR, RNA sequencing, and non-targeted metabolomics. AML12 cells were used for in vitro experiments to validate the findings. Aspirin treatment significantly reduced plasma lipid levels and liver lipid accumulation in HFD-fed mice. RNA sequencing and non-targeted metabolomics identified differentially expressed genes (DEGs) and metabolites (DEMs), respectively. These findings were validated through RT-qPCR for the DEGs and targeted mass spectrometry for the DEMs. Enrichment analyses highlighted several key pathways, including lipid metabolism, PPAR signaling, and bile acid metabolism. Integrated transcriptomic and metabolomic analysis identified 42 overlapped pathways that may mediate the protective effects of aspirin. In vitro experiments confirmed that aspirin reduced lipid accumulation and inflammation in palmitic acid-treated AML12 cells. Molecular docking confirmed strong binding between aspirin/cholic acid and SULT2A3. SULT2A3 was upregulated in MASLD patients and HFD-fed mice. Functional studies revealed SULT2A3 overexpression exacerbated PA-induced lipid accumulation, inflammation, and bile acid dysregulation, whereas its knockdown or aspirin treatment mitigated these effects. Aspirin ameliorates MASLD by modulating SULT2A3-mediated bile acid metabolism and inflammatory pathways. Sult2a3 emerges as a potential target for the treatment of MASLD.