Fig. 2: Inhibition of oxidative stress and the TGF-β/Smad signaling pathway by B. fragilis and the discovery of 1,5-AG by untargeted and targeted metabolomics.
From: The gut microbe Bacteroides fragilis ameliorates renal fibrosis in mice
A Kidney expression of TGF-β/Smad signaling pathway from Sham, UUO and B fragilis-treated UUO mice, assayed by Western blot. B Quantitative analysis of panel A (n = 3). **p = 0.0070 for TGF-β: Sham vs. UUO, p = 0.9998 for TGF-β: Sham vs. UUO + BF, ##p = 0.0076 for TGF-β: UUO vs. UUO + BF; **p = 0.0049 for SMAD2: Sham vs. UUO, p = 0.2267 for SMAD2: Sham vs. UUO + BF, #p = 0.0470 for SMAD2: UUO vs. UUO + BF; *p = 0.0128 for SMAD3: Sham vs. UUO, p = 0.6852 for SMAD3: Sham vs. UUO + BF, #p = 0.0444 for SMAD3: UUO vs. UUO + BF. C Representative Western blot of Nrf2, Keap1, 12-LOX, Rac-1. D Quantitative analysis of panel C (n = 3). *p = 0.0118 for NRF2: Sham vs. UUO, p = 0.4377 for NRF2: Sham vs. UUO + BF, #p = 0.0241 for NRF2: UUO vs. UUO + BF; **p = 0.0065 for KEAP1: Sham vs. UUO, *p = 0.0290 for KEAP1: Sham vs. UUO + BF, #p = 0.0276 for KEAP1: UUO vs. UUO + BF, Comparison in KEAP1 were performed with a two-tailed Student’s t test; **p = 0.0010 for 12-LOX: Sham vs. UUO, p = 0.5931 for 12-LOX: Sham vs. UUO + BF, ##p = 0.0029 for 12-LOX: UUO vs. UUO + BF; **p = 0.0014 for RAC1: Sham vs. UUO, *p = 0.0455 for RAC1: Sham vs. UUO + BF, #p = 0.0351 for RAC1: UUO vs. UUO + BF. E OPLS-DA in the indicated groups (n = 7). F Disturbed metabolic pathways for differential metabolites from sham vs UUO groups. G Heatmap of the differential metabolites. The asterisk indicates that the metabolite can be regulated by B. fragilis. H The concentration of 1,5-AG in serum from Sham, UUO and B. fragilis-treated UUO groups using GC-MS based targeted metabolomics (n = 7). *p = 0.0138 for H: Sham vs. UUO, p = 0.9746 for H: Sham vs. UUO + BF, #p = 0.0317 for H: UUO vs. UUO + BF. I OPLS-DA plot of CKD patients (n = 115) and healthy controls (n = 113) of serum from the Affiliated Hospital of Nanjing University of Chinese Medicine through GC-MS based untargeted metabolomics. J GC-MS based targeted metabolomics of 1,5-AG in the sera of CKD and healthy controls (n = 110) from Ningbo Hospital of Zhejiang University. ***p < 0.0001. K LC-MS based targeted metabolomics of 1,5-AG in the sera of CKD and controls (n = 100) from Putuo People’s Hospital. ***p < 0.0001. Box plots show center line as median, whiskers show maxima and minima, and box limits show upper and lower quartiles. Data are presented as mean ± SD. Comparison in J, K were performed with a two-tailed Mann-Whitney U test. Comparisons in B, D and H were compared using One-Way ANOVA followed by Sidak’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001 (compared with sham or healthy control group), #P < 0.05, ##P < 0.01, ###P < 0.001 (compared with UUO group). Individual data points are independent biological replicates unless otherwise stated.
