Journal: Nature medicine

Article Title: Gut microbiota and intestinal FXR mediate the clinical benefits of metformin

doi: 10.1038/s41591-018-0222-4

Figure Lengend Snippet: a, TR-FRET FXR coactivator recruitment assay to assess whether GUDCA is a FXR antagonist in the presence of CDCA (20 μM). n = 4 replicates/treatment. b, Luciferase activity of control (DMSO), different concentrations of GUDCA or TUDCA without or with FXR agonist CDCA treatment. n = 3 replicates/treatment. P value was determined by one-way ANOVA with Tukey’s correction (F7,16 = 61.69). c, SHP and FGF19 mRNA expression in differentiated Caco-2 cells after treatment with different concentrations of GUDCA or TUDCA with CDCA. n = 3 replicates/treatment. P value was determined by one-way ANOVA with Tukey’s correction (SHP: F7,16 = 4.871, FGF19: F7,16 = 15.41). d, Relative expression of the target gene mRNAs of intestinal FXR in mice treated with vehicle, TCA or TCA and CDCA, GUDCA or TUDCA. n = 5 mice/group. P value was determined by one-way ANOVA with Tukey’s correction (Shp: F4,20 = 38.47, Fgf15: F4,20 = 25.02). e, Relative expression of intestinal Fxr mRNA and its target gene mRNAs in mice treated with metformin on a HFD for 1 week. n = 5 mice/group. P value was determined by two-tailed Student’s t-test (Shp: t8 = 2.779, Fgf15: t8 = 3.383). f, The relative expression of Cyp7a1, Cyp7b1, Cyp8b1 and Cyp27a1 mRNAs in the livers of metformin-treated mice after 1-week HFD feeding. n = 5 mice/group. P value was determined by two-tailed Student’s t-test (Cyp7a1: t8 = 2.805). g, Relative abundance of intestinal Fxr mRNA and its target gene mRNAs in metformin-treated Ampka1fl/fl and Ampka1ΔIE mice on a HFD for 1 week. n = 5 mice/group. P value was determined by one-way ANOVA with Tukey’s correction (Shp: F3,16 = 8.046, Fgf15: F3,16 = 11.83). h, Relative expression of intestinal Fxr mRNA and its target gene mRNAs in antibiotics-treated, microbiota-depleted mice treated with metformin on a HFD for 1 week. n = 5 mice/group. P value was determined by one-way ANOVA with Tukey’s correction (Shp: F3,16 = 6.859, Fgf15: F3,16 = 8.782). i, In microbiota-depleted mice, metformin had no effect in inhibiting the activation of intestinal FXR on top of TCA. The relative expression of Fxr and its target genes in the ileum. n = 6 mice/group. All the data are presented as the mean ± s.e.m.

Article Snippet: After 24 h post-transfection, the cells were exposed to GUDCA (50–200 μM, Sigma-Aldrich, Cat# 06863), or TUDCA (50–200 μM, Sigma-Aldrich, Cat# T0266) in the presence of CDCA (20 μM, Targetmol, Cat# T0847) for 16 h. Luciferase assays were performed using the dual-luciferase assay system (Promega).

Techniques: Luciferase, Activity Assay, Expressing, Two Tailed Test, Activation Assay

Journal: Clinical pharmacology and therapeutics

Article Title: Systems Pharmacology Modeling Predicts Delayed Presentation and Species Differences in Bile Acid-Mediated Troglitazone Hepatotoxicity

doi: 10.1038/clpt.2014.158

Figure Lengend Snippet: Hepatic and systemic disposition of drugs/metabolites are simulated using a physiologically-based pharmacokinetic (PBPK) model ( Drug PBPK Model ). The Bile Acid Homeostasis Model represents hepatobiliary disposition and enterohepatic recirculation of lithocholic acid (LCA) and chenodeoxycholic acid (CDCA) species, and all other (bulk) bile acids. Using bile acid transport inhibition constants of drugs/metabolites (e.g., K i , IC 50 ), altered bile acid disposition is simulated. Increased hepatocellular accumulation of bile acids inhibits hepatic ATP synthesis and decreases intracellular ATP concentrations ( Cellular ATP Model ), leading to necrotic cell death ( Hepatocyte Life Cycle Model ) and elevations in serum biomarkers of hepatocellular injury and function (e.g., ALT, AST, bilirubin) ( Biomarker Model ). Loss of hepatocytes will subsequently influence drug and bile acid disposition (dashed lines), allowing dynamic interaction between kinetics and toxicity mechanisms. Details regarding the construction and structures of sub-models can be found in the .

Article Snippet: In DILIsym, CDCA, LCA, and their conjugates were exclusively modeled as the toxic BAs.

Techniques: Inhibition, Biomarker Discovery

Journal: Clinical pharmacology and therapeutics

Article Title: Systems Pharmacology Modeling Predicts Delayed Presentation and Species Differences in Bile Acid-Mediated Troglitazone Hepatotoxicity

doi: 10.1038/clpt.2014.158

Figure Lengend Snippet: Predicted maximum hepatic accumulation of CDCA and LCA species and DILI responses (i.e., minimum hepatic ATP, minimum viable liver mass, maximum serum ALT) post-dose in human SimPops at oral doses of 200 (green triangle), 400 (blue circle), or 600 (red diamond) mg/day TGZ for 6months (A), and rat SimPops at oral doses of 5 (blue circle) or 25 (red diamond) mg/kg/day for 6months (B).

Article Snippet: In DILIsym, CDCA, LCA, and their conjugates were exclusively modeled as the toxic BAs.

Techniques:

Journal: Clinical pharmacology and therapeutics

Article Title: Systems Pharmacology Modeling Predicts Delayed Presentation and Species Differences in Bile Acid-Mediated Troglitazone Hepatotoxicity

doi: 10.1038/clpt.2014.158

Figure Lengend Snippet: List of parameters varied in the human and rat SimPops and results of multiple regression analysis in human SimPops administered 600 mg/day troglitazone (TGZ) for 6 months Human and rat population samples incorporating variability in parameters governing bile acid homeostasis (Bile Acid SimPops) have been constructed previously. 20 Four parameters in the Drug PBPK Sub-model and two system-specific parameters also were varied. (See supplementary material for methods and data used for construction of SimPops). In the human SimPops administered 600 mg/day TGZ for 6 months, a multiple regression analysis was performed to identify the most important parameters in TGZ-mediated hepatotoxicity using 16 varied parameters as independent variables and minimum hepatic ATP as the dependent variable. Statistical significance and standardized coefficients were calculated using JMP 10.

Article Snippet: In DILIsym, CDCA, LCA, and their conjugates were exclusively modeled as the toxic BAs.

Techniques: Construct, Inhibition