Fig 1: B. adolescentis protects against diet-induced obesity and affects intestinal and circulating ANGPTL4.a, Schematic diagram of B. adolescentis (B.a) supplementation strategy. WD-induced obese C57BL/6 mice were supplemented with drinking water with live (WD + B.a) or heat-killed B. adolescentis (WD + hk-B.a) or saline (WD) for 5 weeks. b, The abundance of B. adolescentis quantified by qPCR using the specific primers in caecum content among three groups. c–k, Parameters measured in the three groups after 5 weeks of treatment: body weight (c). Adipose depot mass including iSAT, eWAT, pWAT and mWAT (d). The localizations and levels of ANGPTL4 in ileum of mice were visualized by immunohistochemistry staining (scale bar, 100 µm) (e). Representative images of each group were shown. ANGPTL4+ area as a percentage of total ileal mucosa area (f). ANGPTL4 mRNA expression levels in the ileum (g). Relative intestinal luminal lipase activity (h). TG levels in the ileum (i). Faecal TG levels (j). Serum ANGPTL4 levels (k). n = 8 biological replicates for each group (b–d,g–k). Data were reproduced in three independent experiments. Data are presented as mean ± s.e.m. Significance was determined by one-way ANOVA (normally distributed) followed by Tukey’s post hoc test or Kruskal–Wallis test (non-normally distributed) followed by Dunn’s test. *P = 0.04 (c), 0.02, 0.04, 0.03 (d) and 0.05 and 0.05 (k). **P = 0.001, 0.004 (b), 0.003, 0.004 and 0.002 (d), 0.001 (f), 0.009 (h), 0.002 (i) and 0.003 and 0.003 (j). ***P < 0.001.Source data
Fig 2: Baseline abundances of two primary degrading microbiota of RS and their relationship with clinical outcomes.(a-d) The two-tailed Pearson correlation between (a) the baseline abundances of B. adolescentis and R. bromii. (b) the baseline abundances of B. adolescentis and the abundance change of R. bromii. (c) the baseline abundances of R. bromii and the abundance change of B. adolescentis. (d) the abundance changes of B. adolescentis and R. bromii. The changes in B. adolescentis and R. bromii are calculated by the log2 fold change after treatment. Baseline abundance values have undergone centred log-ratio (CLR) transformation before correlation analysis. The lines and grey zones show the fitted linear regression lines with 95% confidence intervals. (e) The relationship between the presence of key species (B. adolescentis and R. bromii) and the change of key outcomes after RS treatment. The changes of key outcomes were presented by the log2 fold change after treatment. The violin plots illustrate the kernel density estimation of the data distribution, and the embedded box plots indicate the median and interquartile range. Whiskers extend to 1.5 times the IQR. n = 16 for R. bromii positive individuals, n = 11 for R. bromii negative individuals, n = 9 for B. adolescentis positive individuals, and n = 18 for B. adolescentis negative individuals. Statistical differences between the positive and negative groups were assessed using the two-sided Wilcoxon rank-sum test, with P values displayed above the plots. * P = 0.033 and 0.046 for fat mass and ANGPTL4 (e). The colour indicates the presence (positive, orange) or absence (negative, cyan) of key species. Source data
Fig 3: Effects of RS in germ-free mice with and without B. adolescentis.a, Schematic diagram of B. adolescentis (B.a) supplementation strategy. Orally inoculating B. adolescentis or PBS into germ-free (GF) mice on diets with 20% protein, 45% fat and 35% carbohydrate sourced (from 20% CS or 20% RS and the remaining 80% from maltodextrin) for 8 weeks. b, Abundance of B. adolescentis quantified as total plate count in caecum content. c.f.u., colony-forming unit. c, Disembowelled body weight. d, Glucose excursion curves of intraperitoneal glucose tolerance tests (GTTs). e, Glucose excursion curves of intraperitoneal insulin tolerance tests (ITTs). f, Body mass composition. g, After an 8-week B. adolescentis supplementation, in vivo gut permeability was determined by measurement of serum concentrations of DX-4000-FITC at 1 h after oral gavage. h, Expression of ZO-1 and occludin in the ileum. i, Expression of inflammatory genes in mWAT. j, mRNA expression levels of ANGPTL4 in the ileum. k, Relative intestinal luminal lipase activity. l, TG levels in the ileum. m, Faecal TG levels. Data are mean ± s.e.m. (n = 5 biological replicates per group). *P = 0.04 and 0.03 (d), 0.02 (e), 0.05, 0.04 (f) and 0.03 (k), **P = 0.002 (c), 0.002 (e), and 0.004 (l), ***P < 0.001 based on one-way ANOVA (normally distributed) followed by Dunnett’s test or Kruskal–Wallis test (non-normally distributed) followed by Dunn’s test. * indicates the comparison between RS + B.a and RS + PBS (d,e).Source data
Fig 4: Alleviation of obesity after the 8-week RS intervention in individuals with excess body weight.a, Diagram of the clinical trial. After enrolment, randomization and run-in period, participants consumed either RS or CS alternately and separated by a washout period. During the whole trial, all participants were provided with identical diets. The assessments at each visit are displayed in the diagram. b–d, RS intervention significantly reduced body weight (b), fat mass (c) and waist circumference (d). e,f, Change of VFA and SFA evaluated by MRI. g, Representative abdominal MRI of participants before (left) and after (right) the 8-week RS intervention. Raw (top) and marked (bottom) MRI at navel level. Yellow represents SFA and red represents VFA. h, Change of GIR evaluated by hyperinsulinemic–euglycemic clamp. i, Change of serum TNFα levels. j, Change of serum IL-1β levels. k, Daily faecal lipid excretion, including NEFA, TG and TC after the 8-week interventions with RS or CS. l, Change of serum ANGPTL4 levels. m, Change of serum FGF21 levels. n = 37 individuals (b–d,j,l,m), n = 36 individuals (e,f,i), n = 35 individuals (h) and n = 17 individuals (k) for either RS or CS. Analysis of covariance (ANCOVA) adjusted by baseline value was used for comparison between RS and CS at each visit (b–d). Data are shown as mean (95% confidence interval (CI)). ***P < 0.001. Data are shown as median with IQR (k). Nonparametric Wilcoxon rank-sum test was used to evaluate the significance between the two interventions. ***P < 0.001. Data are shown as box-and-whisker plots (e,f,h–j,l,m). Box plot, median and quartiles; whiskers, data range. *P = 0.025, 0.014 and 0.046 (h–j), **P = 0.004 and 0.002 (f,m), ***P < 0.001 for the between-group difference assessed by the linear mixed model adjusted for intervention order. ††P = 0.003 and 0.002 (h,l). †††P < 0.001 for the within-group change by mixed linear model adjusted for intervention followed by Bonferroni’s test.
Fig 5: RS-influenced gut microbiota restores gut barrier and reduces lipid absorption.Mice were grouped and treated as in Fig. 4 (n = 16 per group). a, Serum levels of inflammatory cytokines in mice colonized with microbiota from RS or CS donors. b, The expression of inflammatory genes in mWAT and in mice colonized with microbiota from RS or CS donors. c, Gut permeability in vivo. d, The expression of ZO-1 and occludin in the ileum. e, The localizations and levels of ZO-1 (red) and occludin (green) in intestinal villus were visualized by immunofluorescence and counterstaining with 4,6-diamidino-2-phenylindole (DAPI) (blue). Representative images of each group are shown (scale bar, 50 µm). f, Quantitative analysis of the positive stained area of ZO-1 and occludin was performed by ImageJ software and calculated as the percentage of total lesion area. g,h, LPS levels in mWAT and in circulation (serum). i, Expression levels of ANGPTL4 in the ileum. j, Relative intestinal luminal lipase activity. k, TG levels in the ileum. l, Faecal TG levels. m, Serum ANGPTL4 levels. Data were reproduced in three independent experiments. Data are presented as mean ± s.e.m. Significance was determined by unpaired two-tailed Student’s t-test (normally distributed) or nonparametric two-sided Wilcoxon rank-sum test (non-normally distributed). **P = 0.005 (a) and 0.001 and 0.010 (b), ***P < 0.001.Source data
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