Fig 1.
Experimental design for disease progression analysis (A) and mRNA sequencing (B). ALT indicates alanine transaminase; AMLN, Amylin liver NASH; AST, aspartate transaminase; MRI, magnetic resonance imaging; NASH, nonalcoholic steatohepatitis; OCA, obeticholic acid; RNA, ribonucleic acid.
Fig 2.
Representative images of liver histopathologic assessments of steatosis (H&E staining), inflammation (galectin-3 staining), and fibrosis (Col1a1 staining).
Col1a1 indicates collagen type 1 alpha 1 chain; H&E, hematoxylin and eosin; OCA, obeticholic acid. Stains are magnified 20×, scale bar = 100 μm.
Fig 3.
Levels of histopathology markers quantified by percentage fractional area for steatosis (A), inflammation (B), and fibrosis (C). Data are percentage of fractional area change from baseline with minimum, maximum, and median values and interquartile range (Q1 and Q3). *p < 0.05, ***p < 0.001, ****p < 0.0001 compared with vehicle by 1-way analysis of variance with Dunnett post hoc test. #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001 by 1-way analysis of variance with Bonferroni post hoc test. Col1a1 indicates collagen type 1 alpha 1 chain; OCA indicates obeticholic acid. Vehicle, n = 10; OCA 45/30 mg/kg, n = 11; OCA 60/30 mg/kg, n = 11; INT-787 10 mg/kg, n = 11; INT-787 30 mg/kg, n = 10; INT-787 60 mg/kg, n = 10; INT-787 120 mg/kg, n = 11.
Fig 4.
Levels of the plasma liver enzymes ALT (A) and AST (B) at termination. Data are individual plasma ALT and AST (U/L) values; minimum, maximum, and median values; and interquartile range (Q1 and Q3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared with vehicle by 1-way analysis of variance with Dunnett post hoc test. #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001 by 1-way analysis of variance with Bonferroni post hoc test. ALT indicates alanine transaminase; AST, aspartate transaminase; OCA, obeticholic acid. Vehicle, n = 10; OCA 45/30 mg/kg, n = 11; OCA 60/30 mg/kg, n = 11; INT-787 10 mg/kg, n = 11; INT-787 30 mg/kg, n = 10; INT-787 60 mg/kg, n = 9; INT-787 120 mg/kg, n = 11 for animals with plasma ALT and AST measurements at termination.
Fig 5.
Body weight (g) at baseline (A) and end of study (week 8) (B). Data are individual body weights; minimum, maximum, and median values; and interquartile range (Q1 and Q3). OCA indicates obeticholic acid. Vehicle, n = 9; OCA 45/30 mg/kg, n = 10; OCA 60/30 mg/kg, n = 11; INT-787 10 mg/kg, n = 11; INT-787 30 mg/kg, n = 10; INT-787 60 mg/kg, n = 9; INT-787 120 mg/kg, n = 11 for animals with body weight measurements at day 57.
Fig 6.
Genes associated with farnesoid X receptor signaling in the liver significantly up- or downregulated after treatment with OCA 30 mg/kg or INT-787 30 mg/kg compared with vehicle.
Differences in expression levels of genes meeting the adjusted p < 0.05 threshold for significance are shown in red or green. Gray circles represent genes in the farnesoid X receptor signaling pathway that were not significantly up- or downregulated after treatment. OCA indicates obeticholic acid; padj, adjusted p value. Vehicle, n = 10; OCA 30 mg/kg, n = 9; INT-787 30 mg/kg, n = 9.
Fig 7.
Genes associated with lipid metabolism in the liver significantly up- or downregulated after treatment with OCA 30 mg/kg or INT-787 30 mg/kg compared with vehicle.
Differences in expression levels of genes meeting the adjusted p < 0.05 threshold for significance are shown in red or green. Gray circles represent genes in the lipid metabolism pathway that were not significantly up- or downregulated after treatment. OCA indicates obeticholic acid; padj, adjusted p value. Vehicle, n = 10; OCA 30 mg/kg, n = 9; INT-787 30 mg/kg, n = 9.
Fig 8.
Genes associated with stellate cell activation (fibrosis) in the liver significantly up- or downregulated after treatment with OCA 30 mg/kg or INT-787 30 mg/kg compared with vehicle.
Differences in expression levels of genes meeting the adjusted p < 0.05 threshold for significance are shown in red or green. Gray circles represent genes in the stellate cell activation pathway that were not significantly up- or downregulated after treatment. OCA indicates obeticholic acid; padj, adjusted p value. Vehicle, n = 10; OCA 30 mg/kg, n = 9; INT-787 30 mg/kg, n = 9.