Fig. 2: B. thetaiotaomicron or B. fragilis leveraged on sphingolipids biosynthesis to hinder NAC permeability. | Nature Communications

Fig. 2: B. thetaiotaomicron or B. fragilis leveraged on sphingolipids biosynthesis to hinder NAC permeability.

From: Intestinal permeability of N-acetylcysteine is driven by gut microbiota-dependent cysteine palmitoylation

Fig. 2

a Heat-map analysis of top 21 metabolites in intestinal fluid-derived metabolites between NAC-perfused GF and SPF rats. Each column represents one independent sample, and each row represents one metabolite. The color indicates the relative abundance of metabolites in each group. b Volcano plots to illustrate metabolites difference between NAC-perfused GF and SPF rats. Dots corresponding to significant lipids (P < 0.05, Student’s t tests) were colored, in which lipids with increased fold change were colored as red, and sphingolipids with decreased fold change pertained to green. c Histogram presentation of the KEGG pathway. A total of 99 differentiated functional pathways were successfully annotated and grouped into 10 functional categories. P values were determined using two-sided Fisher’s exact tests with Benjamini-Hochberg correction for multiple testing. d Summary of genus and species taxonomic changes in the gut microbiome of SPF rats following perfusion with 30 mg/L NAC. Taxonomic cladogram (e) and histogram (f) were generated by LEfSe of metagenomic analysis data in intestinal fluid samples derived from NAC-perfused and NAC-unperfused segments of SPF rats. g RT-qPCR to determine the 16S rRNA of B. thetaiotaomicron (left coordinate) and B. fragilis (right coordinate) absolute abundance in each group. n  =  6 per group. Data are the means ± SD, *P < 0.05, **P < 0.01 (Student’s t tests). h Correlation heat-map of microbial species abundance and metabolites after NAC perfusion in the intestine. Statistical significance was assessed using two-sided Pearson correlation analyses, with *P < 0.05, **P < 0.01, ***P < 0.001 indicating significant differences. i Cysteine-SL de novo synthesis pathway. SPT (serine palmitoyl transferase), KDSR (3-keto-dihydrosphignosine reductase), DES (desaturase), CerS (ceramide synthase), CDase (ceramidases). j Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was utilized to identify and quantify palmitoyl-CoA (left panel) and cysteine-3-ketosphinganine (middle panel). The curve of left panel is representative image of each group. Data of right panel are presented as mean ± SD. Statistical significance was assessed using one-way ANOVA, with *P < 0.05, **P < 0.01 indicating significant differences. Source data are provided as a Source Data file.

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