Multi-biotics supplementation is associated with partial attenuation of CDAHFD-induced steatotic liver disease in mice: a multi-omics analysis

Gut microbiota plays an integral role in steatotic liver disease (SLD) and steatohepatitis (SH). We evaluated whether Multi-biotics supplementation (a freeze-dried product containing prebiotics, probiotics, and postbiotics) was associated with differences in diet-induced steatotic liver injury. 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 (SLD model, n = 24) and 30 (SH-HCC, n = 24). Mice in both CDAHFD-fed groups developed steatotic liver injury. At 8 weeks, the CDAHFD+Multi-biotics group had lower semiquantitative steatosis and inflammation scores than the CDAHFD group, whereas quantitative ORO area was not significantly different. At 30 weeks, both the steatosis score and ORO-positive area were lower in the CDAHFD+Multi-biotics group. 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 identified differences in selected hepatic transcripts, including higher Nr4a1 expression in the CDAHFD+Multi-biotics group. In separate qRT-PCR and Western blot analyses selected inflammatory transcripts and the total protein abundance of TLR4, TGF-β1, and NF-κB p65 were lower than those in the CDAHFD group. Multi-biotics supplementation was associated with modulation of gut microbiota and with partial improvements in hepatic inflammation and steatosis in this model. Because the design was preventive and the findings are correlative, causal experiments are required before mechanistic or therapeutic conclusions can be drawn.