Akkermansia muciniphila inversely correlates with the onset of inflammation, altered adipose tissue metabolism and metabolic disorders during obesity in mice

Marc Schneeberger, Amandine Everard, Alicia G Gómez-Valadés, Sébastien Matamoros, Sara Ramírez, Nathalie M Delzenne, Ramon Gomis, Marc Claret, Patrice D Cani, Marc Schneeberger, Amandine Everard, Alicia G Gómez-Valadés, Sébastien Matamoros, Sara Ramírez, Nathalie M Delzenne, Ramon Gomis, Marc Claret, Patrice D Cani

Abstract

Recent evidence indicates that the gut microbiota plays a key role in the pathophysiology of obesity. Indeed, diet-induced obesity (DIO) has been associated to substantial changes in gut microbiota composition in rodent models. In the context of obesity, enhanced adiposity is accompanied by low-grade inflammation of this tissue but the exact link with gut microbial community remains unknown. In this report, we studied the consequences of high-fat diet (HFD) administration on metabolic parameters and gut microbiota composition over different periods of time. We found that Akkermansia muciniphila abundance was strongly and negatively affected by age and HFD feeding and to a lower extend Bilophila wadsworthia was the only taxa following an opposite trend. Different approaches, including multifactorial analysis, showed that these changes in Akkermansia muciniphila were robustly correlated with the expression of lipid metabolism and inflammation markers in adipose tissue, as well as several circulating parameters (i.e., glucose, insulin, triglycerides, leptin) from DIO mice. Thus, our data shows the existence of a link between gut Akkermansia muciniphila abundance and adipose tissue homeostasis on the onset of obesity, thus reinforcing the beneficial role of this bacterium on metabolism.

Figures

Figure 1. Kinetic evolution of body weight…
Figure 1. Kinetic evolution of body weight and adiposity following chronic HFD administration.
(A) Body weight gain (g) and (B) adiposity (% of eWAT on total body weight) measured after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a control diet (CT) (n = 6/group). Data are presented as the mean ± SEM. Data are significantly different (P < 0.05) according to the unpaired two-tailed Student t-test. *indicates a significant difference versus CT (P < 0.05).
Figure 2. Time series evolution of markers…
Figure 2. Time series evolution of markers of inflammation and macrophage infiltration in white adipose tissue following HFD administration.
mRNA expression of (A) Tnf (encoding TNF-α), (B) Ccl2 (encoding MCP-1), (C) Itgax encoding CD11c), (D) Emr1 (encoding F4/80), (E) Lbp (encoding LBP), (F) Il6 (encoding Il-6) and (G) Il1b (encoding IL-1β) measured in the adipose tissue after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a control diet (CT) (n = 6/group). Data are presented as the mean ± SEM. Data are significantly different (P < 0.05) according to the unpaired two-tailed Student t-test. *indicates a significant difference versus CT (P < 0.05).
Figure 3. Time series evolution of markers…
Figure 3. Time series evolution of markers of fatty acid oxidation and browning in white adipose tissue following HFD treatment.
mRNA expression of (A) Pppargc1a (encoding PGC1-α), (B) Pppargc1b (encoding PGC1-β), (C) Cpt1a (encoding CPT-1a), (D) Acacb (encoding ACC2), (E) Cidea (encoding CIDEA), (F) Elovl3 (encoding ELOVL3), (G) Prdm16 (encoding PRDM16) and (H) Acox1 (encoding ACOX1) measured in the adipose tissue after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a control diet (CT) (n = 6/group). Data are presented as the mean ± SEM. Data are significantly different (P < 0.05) according to the unpaired two-tailed Student t-test. *indicates a significant difference versus CT (P < 0.05).
Figure 4. Time series evolution of markers…
Figure 4. Time series evolution of markers of lipogenesis and adipogenesis in white adipose tissue following HFD treatment.
mRNA expression of (A) Dgat2 (encoding DGAT2), (B) Acaca (encoding ACC1), (C) Fasn (encoding FAS), (D) Pparg (encoding PPAR-γ) and (E) Cebpa (encoding CEBP-α) measured in the adipose tissue after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a control diet (CT) (n = 6/group). Data are presented as the mean ± SEM. Data are significantly different (P < 0.05) according to the unpaired two-tailed Student t-test. *indicates a significant difference versus CT (P < 0.05).
Figure 5. Time series evolution of orexigenic…
Figure 5. Time series evolution of orexigenic and anorexigenic markers in the hypothalamus and circulating levels of metabolic parameters following HFD treatment.
mRNA expression of (A) AgRP (encoding AGRP), (B) Npy (encoding NPY), (C) Pomc (encoding POMC) measured in the hypothalamus,. Circulating levels of (D) glucose, (E) triglycerides, (F) leptin and (G) insulin measured in the serum of mice after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a control diet (CT) (n = 6/group). Data are presented as the mean ± SEM. Data are significantly different (P < 0.05) according to the unpaired two-tailed Student t-test. *indicates a significant difference versus CT (P < 0.05).
Figure 6. Time series evolution of specific…
Figure 6. Time series evolution of specific gut bacteria following HFD treatment.
Quantification of (A) Akkermansia muciniphila, (B) Bifidobacterium spp., (C) Roseburia spp. (D) Lactobacillus spp., (E) Bacteroides/Prevotella spp, (F) Bilophila wadsworthia and (G) total gut bacteria abundance after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a control diet (CT) and expressed as Log cells per g of caecal content (n = 6/group). Data are presented as the mean ± SEM. Data are significantly different (P < 0.05) according to the unpaired two-tailed Student t-test. *indicates a significant difference versus CT (P < 0.05).
Figure 7. Individuals factor map.
Figure 7. Individuals factor map.
The individual factor map presents the repartition of the samples (dots) in the multifactorial analysis’ plane. Time and diet (squares) are presented as illustrative (inactive) qualitative factors. Samples are colored according to treatment’s duration. Circles regroup all samples from a specific diet for a single time point, except week 3 where all samples are grouped together.
Figure 8. Hierarchical clustering.
Figure 8. Hierarchical clustering.
Hierarchical clustering of the samples on the principal components of the multifactorial analysis. This graph illustrates the similarities and the clustering between different samples. The smaller the distance linking samples is (resulting from the addition of horizontal distances in dark line), the more similar these samples are for the parameters measured.
Figure 9. Correlation circle and heat map…
Figure 9. Correlation circle and heat map correlation showing associations between bacterial taxa and adipose tissue metabolic parameters.
Multifactorial analysis (A) correlation map and (B) heat map of the Spearman r correlations between the bacterial genera and the metabolic parameters measured in the adipose tissue of mice after 3 weeks (3w); 6 weeks (6w); 12 weeks (12w) and 16 weeks (16w) of a high-fat diet (HFD) or a CT diet (CT) (n = 6/group). Data are presented as the mean ± SEM. Squared cells depict significance following the Spearman correlation and FDR correction for multiple comparisons *P < 0.05.

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