Bridging the Gut and Liver: Multi-biotics modulate microbiota composition and corroborate transcriptomic improvements in MASLD

Background Gut microbiota play an integral role in metabolic dysfunction-associated steatotic liver disease (MASLD). We aimed to clarify the potential ameliorative effects of Multi-biotics (a freeze-dried product containing prebiotics, probiotics, and postbiotics) on diet-induced MASLD. Methods Forty-eight C57BL/6 mice were evenly divided into three groups according to diet type: standard diet (SD); choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD); and CDAHFD supplemented with Multi-biotics (CDAHFD+Multi-biotics). The blood, feces, and liver tissues were collected at weeks 8 (early stage MASLD, n = 24) and 30 (late-stage MASLD, n = 24). Results Mice in the two CDAHFD groups developed steatohepatitis at weeks 8 and 30, unlike those in the SD group. The NAFLD activity score showed that the addition of Multi-biotics was significantly associated with reduced steatosis at weeks 8 and 30. Gut microbiota, including the Muribaculaceae and Ruminococcaceae families, were significantly decreased in the CDAHFD group, compared with those in the SD and CDAHFD+Multi-biotics groups at week 8. Transcriptomic analysis indicated that mice in the CDAHFD+Multi-biotics group showed upregulated macrophage-targeting anti-inflammatory signals, such as Nr4a1 , whereas the pro-inflammatory TLR4 signal was downregulated. These transcriptomic findings were corroborated by qPCR and Western blot analysis, which showed consistent trends at the mRNA and protein levels, respectively. Conclusions Multi-biotics may modulate gut microbiota and support improvements in hepatic inflammation and steatosis. The integration of multi-omics data provides valuable mechanistic insight into the therapeutic potential of Multi-biotics.