Fig. 7: The anti-fibrotic effects of B fragilis, 1,5-AG and Mad in the adenine model.

A Representative photomicrographs of the H&E staining and Masson’s trichrome staining from left kidneys of sham, adenine, B. fragilis-treated UUO mice, 1,5-AG-treated adenine mice and Mad-treated adenine mice (H&E and Masson’s staining; scale bar, 100 μm, magnification, ×200). B Bar graphs depict renal injury scores and renal interstitial fibrosis scores based on H&E staining or Masson’s trichrome staining (n = 5). ***p  < 0.0001 for injury score: Control vs. Ade, ^##p = 0.0045 for injury score: Ade vs. BF, ^#p = 0.0421 for injury score: Ade vs. 1,5-AG, ^###p = 0.0001 for injury score: Ade vs. Mad; ***p  < 0.0001 for fibrosis score: Control vs. Ade, ^###p  < 0.0001 for fibrosis score: Ade vs. BF, ^###p = 0.0002 for fibrosis score: Ade vs. 1,5-AG, ^###p < 0.0001 for fibrosis score: Ade vs. Mad. (C) Biochemical parameters including blood urea nitrogen (BUN), serum creatinine (Scr) (n=9). ***p  < 0.0001 for BUN: Control vs. Ade, ^##p = 0.0029 for BUN: Ade vs. BF, ^###p = 0.0009 for BUN^: Ade vs. 1,5-AG, ^###p < 0.0001 for BUN: Ade vs. Mad; ***p  < 0.0001 for SCR: Control vs. Ade, ^###p  < 0.0001 for SCR: Ade vs. BF, ^###p  < 0.0001 for SCR: Ade vs. 1,5-AG, ^###p ⁼ 0.0002 for SCR: Ade vs. Mad. (D) Kidney expression of FN, Col I and α-SMA from Sham, UUO and B. fragilis-treated adenine mice, assayed by Western blot. E Quantitative analysis of Fig. 7D (n = 3). *p = 0.0166 for FN: Control vs. Ade, ^##p = 0.0080 for FN: Ade vs. BF, Comparison in FN were performed with a two-tailed Student’s t test^; ***p  < 0.0001 for COL-1: Control vs. Ade, *p = 0.0335 for COL-1: Control vs. BF, ^###p  < 0.0001 for COL-1: Ade vs. BF; ***p  < 0.0001 for α-SMA: Control vs. Ade, **p = 0.0082 for α-SMA: Control vs. BF, ^###p = 0.0002 for α-SMA: Ade vs. BF. F Kidney expression of FN, Col I and α-SMA from sham, adenine and 1,5-AG-treated adenine mice, assayed by Western blot. G Quantitative analysis of Fig. 7F (n = 3). ***p = 0.0010 for FN: Control vs. Ade, p = 0.0822 for FN: Control vs. 1,5-AG, ^#p = 0.0128 for FN: Ade vs. 1,5-AG; *p = 0.0108 for COL-1: Control vs. Ade, ^#p = 0.0266 for COL-1: Ade vs. 1,5-AG, Comparison in COL^-1 were performed with a two-tailed Student’s t test; **p = 0.0040 for α-SMA^: Control vs. Ade, p = 0.8409 for α-SMA: Control vs. 1,5-AG, ^##p = 0.0085 for α-SMA: Ade vs. 1,5-AG. (H) Kidney expression of FN, Col I and α-SMA from control, adenine and Mad-treated adenine mice, assayed by Western blot. I Quantitative analysis of Fig. 7H (n=3). *p = 0.0191 for FN: Control vs. Ade, p = 0.5363 for FN: Control vs. Mad, ^##p = 0.0048 for FN: Ade vs. Mad; **p = 0.0018 for COL-1: Control vs. Ade, p = 0.9225 for COL-1: Control vs. Mad, ^##p = 0.0030 for COL-1: Ade vs. Mad; **p = 0.0044 for α-SMA: Control vs. Ade, p = 0.4676 for α-SMA: Control vs. Mad, ^#p = 0.0198 for ^α-SMA: Ade vs. Mad. J Western blot shows SGLT2 protein expression in kidneys from control, adenine and adenine + BF groups. K Quantitative analysis of Fig. 7J (n = 3). **p = 0.0078 for K: Control vs. Ade, *p = 0.0428 for K: Control vs. BF, ^#p = 0.0238 for K: Ade vs. BF, Comparison in K were performed with a two-tailed Student’s t test. Data are presented as mean ± SD. Comparisons in B, C, E, G and I were compared using One-Way ANOVA followed by Sidak’s multiple comparisons test. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 (compared with control group), ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 (compared with adenine group). Individual data points are independent biological replicates unless otherwise stated.