Figure 5.

DR modulates composition of intestinal flora. Two-month-old mice were fed with AL diet or 30% DR diet for 14 days before MTX administration and the diet regimen was continued afterward. Then, mice were intraperitoneally injected with saline as control or MTX for 2 days at a dose of 120mg/kg (d-1) and 60mg/kg (d0) Fecal samples were randomly collected from mice before and on day 2 after MTX injection for 16S rRNA gene sequencing (n = 8–10 mice per group randomly picked from 2 independent experiments) and qPCR analysis (n = 5 mice per group randomly picked from 2 independent experiments). (A) Variation of intestinal flora structure of indicated groups along PC1 and PC2 of PCoA based on the Bray–Curtis distance. (B) Relative abundance of the intestinal flora of indicated groups on the Order level showed by 16S rRNA gene sequencing. (C) LDA scores in the fecal microbiomes of indicated groups. LDA score>2 and top nine bacteria were shown. (D) Relative abundance of the Lactobacillus showed by 16S rRNA gene sequencing. (E) qPCR analysis of the amounts of total bacteria and Lactobacillus of indicated groups (n = 5 mice per group, randomly picked from 2 independent experiments). Results were displayed as mean±SD. n.s; nonsignificant; *: P < .05; **: P < .01 by two-tailed, Student’s t test. AL: mice on AL diet; DR: mice on DR diet; AL+MTX: mice on AL diet and received MTX injection; DR+MTX: mice on DR diet and received MTX injection; d-1: day −1 before MTX injection; d2: day 2 after MTX injection.