Unveiling the role of sex in the metabolism of indoxyl sulfate and apixaban

Figures

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  Fig. 1. Indoxyl Sulfate levels in mice plasma quantified by HPLC (A), Apixaban levels in mice plasma quantified by LC-MS (B). WT Apixaban-treated females [control (KCl) and IS pooled] in orange, WT Apixaban-treated males WT [control (KCl) and IS pooled] in lilac. Grouped comparisons were made to assess sex related differences in Apixaban plasma levels regardless of treatment. Bleeding time (C). Pairwise comparisons were performed using Mann-Whitney U test, shown in black treatment differences within groups and in green sex related differences. Data are shown as median with interquartile range *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (n = 6–20).

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  Fig. 2. Sex-dependent differences in mRNA Expression of efflux transporters and metabolic enzymes of apixaban in mice ileum (A), liver (B) and kidney (C) of apixaban-treatment mice. Pairwise comparisons of pooled—Males vs. Females—were performed using Mann-Whitney U test. Data are shown as median with interquartile range *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (n = 12–20).

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  Fig. 3. Scatterplot of IS and Apixaban levels in males (A) and females (C). Spearman correlation coefficient was applied to assess linear correlation between Apixaban and IS levels. Cyp3a11 mRNA Expression levels in male (B) and female (D) liver. Pairwise comparisons were performed using Mann-Whitney U test. Data are shown as median with interquartile range *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (n = 6–12).

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  Fig. 4. mRNA Expression of Scl22a6 and Scl22a8 in mice kidney. Mice were pooled regarding treatment. Pairwise comparisons—Males vs. Females—were performed using Mann-Whitney U test. Data shown are as median with interquartile range *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (n = 13–20).

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  Fig. 5. Sex-dependent differences in mRNA Expression of efflux transporters and metabolic enzymes in mice ileum (A), liver (B) and kidney (C) involved in IS metabolism. Samples with IS or control treatment were pooled by sex. Pairwise comparisons—Males vs. Females—were performed using Mann-Whitney U test (green). Data shown as median with interquartile range *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 (n = 11–17).

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  Fig. 6. Differences in mRNA Expression of efflux transporters (A) and metabolic enzymes (B) in liver of Wild-type Vs. AhR−/− mice. For Cyp1a1, Cyp1a2 and Cyp2e1 males and females were pooled. Pairwise comparisons were performed using Mann-Whitney U test (blue). For Abcg2 and Sult1a1, Kruskal Wallis statistical test followed by Dunn’s test was used for sex related comparisons (green). Pairwise comparisons were performed using Mann-Whitney U test (black). Data shown as median with interquartile range *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 (n = 5—17).

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  Fig. 7. Protein expression changes in the mouse liver. PCA of global expression changes in mice liver (A). Sex is the main clustering variable. Volcano plots of differentially expressed proteins (blue) (B–E), Table with the number of differentially expressed proteins across experimental groups (F), Heatmap containing the pathway enrichment analysis results (G) (Downregulated pathways in blue and upregulated in red).

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  Fig. 8. Protein expression of efflux transporters and metabolism enzymes related with IS or Apixaban metabolism. Heatmap (A) Downregulated proteins appear colored in blue and upregulated in red. Table containing logFC, t, P.value and FDR (B) for the proteins of interest amongst the different group comparison.