Intestinal neutral ceramidase deficiency prevents intestinal barrier dysfunction and enhances gut fucosylation in mice with MASH. (IMAGE)

First Hospital of Jilin University

Intestinal neutral ceramidase deficiency prevents intestinal barrier dysfunction and enhances gut fucosylation in mice with MASH.

Caption

WT mice or Asah2ΔIEC mice were fed the WD for 12 months. (A) The levels of LBP in serum, lung and liver tissues. (B) The level of serum LPS in the portal veins. (C) Plasma levels of dextran-FITC for evaluating gut permeability. (D) Alcian blue-PAS staining of colonic tissue. Arrows indicate the PAS staining of glycans. (E) Quantification of goblet cells (left) and glycan-containing cells (right) in the crypt. (F) Representative enrichment plots for the hallmark tight junction pathway and mucin-type O glycan biosynthesis pathway. The black vertical line at the bottom shows where the members of the gene set appear in the ranked list of genes. The graded red to blue bars on the x axis represent the DESeq2 statistic values (Asah2ΔIEC mice vs WT), with red and blue denoting upregulation and downregulation, respectively. (G) Heatmap of genes involved in the fucosylation pathway and tight junction pathway in the colon. (H) Real-time PCR analysis of the mRNA levels of indicated genes encoding fucosylation and tight junction proteins in the colon. (I) UEA-1 immunohistochemistry staining of colonic tissue with quantification of UEA-1-positive area. (J) Immunofluorescent staining of mucin 2 (Muc2, green) and UEA-1 (red) of LI and SI and quantification of Mucin 2+UEA-1+ cells per crypt. Statistical comparisons were performed using two-tailed unpaired t-test (A–C, E, I) and two-way ANOVA with Sidak’s multiple comparisons test (H). n=5. Independent biological samples. *p<0.05, ** p<0.01, *** p<0.001. Errors bBars represent mean±SEM. Scale bar represents 100 µm. ANOVA, analysis of variance; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate; Fut, fucosyltransferases; IEC, intestinal epithelial cell; LBP, LPS-binding protein; LI, larger intestine; LPS, lipopolysaccharide; MASH, metabolic dysfunction-associated steatohepatitis; mRNA, messenger RNA; ns, not significant; PAS, periodic acid-Schiff; SI, small intestine; TNFα, tumour necrosis factor-alpha; WD, Western diet; WT, wild-type.

Credit

By Ting Wang, Liang Chen, Chao Lei, et al.

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