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Fig. 6 | Microbiome

Fig. 6

From: Estrogen-mediated gut microbiome alterations influence sexual dimorphism in metabolic syndrome in mice

Fig. 6

Intestinal alkaline phosphatase (IAP) mediates sexual dimorphism in gut microbiota in the context of MS. a, b mRNA expression of two major IAP isozymes AKP3 and AKP6 in the duodenum of five groups (male, female, male+E2, OVX, and OVX+E2; n = 5 per group). c Immunohistochemical (IHC) localization of IAP in the duodenum. The enlarged view of single villus (circled in the main figures) has also been placed under each picture with an arrow head indicating the location of IAP in a circular fashion in the tip of small intestinal villus. d Western blot (WB) analysis showing the protein levels of IAP. e Duodenal tissue total IAP activity. f WB analysis of IAP using cell lysates collected from differentiated Caco-2 cell with enterocytes-like features after stimulating them (in vitro) with 17β-estradiol (10 nM) in a time-dependent manner. g AKP3 mRNA expression of male (M) and male +ISO (M+ISO) groups. h, i Representative IHC staining pictures showing the IAP expression. j WB analysis showing the protein levels of IAP. k Total IAP activity. l In vitro stimulation of Caco-2 cells with 25 μM concentration of either genistein (G) or daidzein (D) or G+D combination, followed by WB analysis of IAP. m In vitro stimulation of Caco-2 cells in the presence or absence of L-phenylalanine (10 mM), G+D (25 μM), and 17β-E (10 nM), followed by WB analysis of IAP. q-PCR-based quantification of Proteobacteria, Bifidobacterium (B) to Enterobacteriacea (E) ratio and relative abundance of Akkermansia and Firmicutes (FIR) to Bacteroidetes (BAC) ratio (nq), markers of metabolic endotoxemia (serum LPS, LBP and intestinal permeability) (rt), low-grade chronic inflammation (TNF-α) (u), and markers of metabolic syndrome (body weight and GTT) (v, w) in the male (M), female (F), and F+L-phe (10 mM)-treated groups. Data was shown as mean ± SEM. Box plots (box showing the median, and the 25th and 75th percentiles, and the whiskers of the graph show the largest and smallest values) were also used. Data with different superscript letters are significantly different (P < 0.05). Student’s t test or ordinary or repeated measures one-way ANOVA followed by Tukey’s multiple comparisons test

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