Intestinal neutral ceramidase deficiency alters gut microbiota and attenuates MASH. (IMAGE)
Caption
The 16S rRNA gene sequencing was performed to characterise gut microbiota composition in mice fed a WD for 12 months. (A) PCoA of faecal microbiota β diversity based on Jaccard distance. (B) α diversity comparison based on Chao1. (C) Heat map of differential faecal microorganisms for top 30 species. (D) Cladogram depicting LEfSe analysis of differentially abundant bacteria in WT mice and Asah2ΔIEC mice. The green colour indicates enriched taxa in the WT mice and the yellow colour indicates enriched taxa in the Asah2ΔIEC mice. The size of each circle is proportional to the taxon’s abundance. (E) LDA scores of the differentially abundant taxa shown in (D). Taxa enriched in microbiota from Asah2ΔIEC mice (yellow) are indicated with a negative LDA score related to that from WT mice (green). Indicate whether observed changes are statistically significant and describe the test applied. (F–J) Germ-free mice were transplanted with stools from Asah2ΔIEC (GF-Asah2ΔIEC) or Asah2fl/fl (GF-WT) and fed an HSPC diet for 4 months. (F) Schematic of FMT experiment and ratio for liver to body weight. Created with BioRender.com. (G) Levels of serum LPS in the portal vein and serum hydroxyproline. (H) Histological determination of hepatic steatosis by H&E staining with quantification of per cent lipid droplet area and inflammatory cells. Arrows indicate immune cell infiltration and inflammation. The area between circles indicates fibrosis. (I) Level of hepatic TG. (J) Representative images of liver sections stained with Masson’s trichrome. (K) Hepatic levels of Col1a1, Col1a2, Col3a1, Fn1 and Acta2 mRNA. Statistical comparisons were performed using Adonis test (A), Wilcoxon test (B), Kruskal–Wallis test (E), two-tailed unpaired t-test (F–J) and two-way ANOVA with Sidak’s multiple comparisons test (K). n=5 (A–E, WT mice). n=4 (A–E, Asah2ΔIEC mice). n=5 (F–K). Independent biological samples. *p<0.05, ** p<0.01, *** p<0.001. Error bars represent mean±SEM. Scale bar represents 100 µm. ANOVA, analysis of variance; FMT, faecal transplantation of gut microbiota; GF, germ-free; HSPC, hydrogenated vegetable oil, sucrose, palmitate and cholesterol; IEC, intestinal epithelial cell; LDA, linear discriminant analysis; LEfSe, linear discriminant analysis effect size; LPS, lipopolysaccharide; MASH, metabolic dysfunction-associated steatohepatitis; mRNA, messenger; ns, not significant; PCoA, principal coordinates analysis; rRNA, ribosomal RNA; TG, triglyceride; WD, Western diet; WT, wild-type.
Credit
By Ting Wang, Liang Chen, Chao Lei, et al.
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