Fig. 4: Two weeks of protein mix is not enough to affect weight gain and development of insulin resistance in HFHS-fed mice but induces changes in microbial composition, SCFA production, and postprandial metabolite profile. | Nature Communications

Fig. 4: Two weeks of protein mix is not enough to affect weight gain and development of insulin resistance in HFHS-fed mice but induces changes in microbial composition, SCFA production, and postprandial metabolite profile.

From: Feeding diversified protein sources exacerbates hepatic insulin resistance via increased gut microbial branched-chain fatty acids and mTORC1 signaling in obese mice

Fig. 4

a Total body weight gain of mice fed with HFHS-C (purple) or HFHS-PM (purple, hatched) diet during a 2-week study. Mice were food-deprived for 12 h after 2 weeks of dietary treatment and food was given back to animals for 15 min. Blood was collected 30 min later by saphenous vein for postprandial (b) glycemia and (c) insulinemia assessment. (n = 12 biologically independent mice for both groups). Gut microbiota differences are represented by (d) a histogram plotting differentially abundant bacteria detected by LEfSe using a threshold of LDA score >2.5 between HFHS-C (green) and HFHS-PM (blue), (e) quantification of fecal Akkermansia muciniphila evaluated by gene expression through qPCR, and (f) Shannon and (g) Simpson’s reciprocal diversity indexes. n = 6 biologically independent mice for both groups. Mice selected for gut microbiota composition analysis were the closest to the median on main physiological parameters (body weight, glycemia, and insulinemia). Fecal levels of major SCFA (h) acetic, (i) butyric, and (j) propionic acid and minor SCFA (k) valeric, (l) isobutyric, and (m) isovaleric acid after 2 weeks of dietary intervention. Two hours following refeeding after the food deprivation, submandibular blood was collected for plasma metabolite profile determination. Mice were euthanized and muscle and liver were harvested for metabolite profiles. Hepatic (n) organic acid profile and (o) acylcarnitines (AC) profiles. n = 12 biologically independent mice for both groups. Data are represented as means ± s.e.m for panels a–g and median, interquartile range, minimum and maximum for panels h–o. Statistical analyses were performed using a two-tailed Student’s t-test or its nonparametric equivalent Mann-Whitney test. Detailed significant differences are recorded as follows: *p < 0.05, **p < 0.01. Exact p-values for trends (0.05 ≤ p-value < 0.10) are recorded on graphs for an additional indication. See also Supplementary Figs. 5–7 and Supplementary Tables 4–6. Short-chain AC: C2 to C4; Medium-chain AC: C5 to C10; Long-chain AC: C12 to C22.

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