Dynamic Alterations in the Gut Microbiota and Metabolome During the Development of Methionine-choline-deficient Diet-induced Nonalcoholic Steatohepatitis

Aim: To investigate changes in gut microbiota and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.

Methods: Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.

Results: The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with liver fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut microbiota dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic acid, hexadecane, palmitic acid, and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic acid, cholesterol, arachidic acid, tetracosane, and stearic acid were selected.

Conclusion: The MCD diet induced persistent alterations in the gut microbiota and metabolome.