Fig 1.
(A) ALT plasma levels compared to vehicle, with F573 significantly lower (p = 0.01). (B) AST plasma levels compared to vehicle (p = 0.42). Data points show means and SEM, 95% confidence interval (n = 20 per group). Aspartate aminotransferase (AST), Alanine aminotransferase (ALT).
Fig 2.
(A) TUNEL staining of tissue samples, reporting relative percentage of TUNEL nuclei compared to DAPI. Significantly lower TUNEL staining was found in the F573 and BMX-001 groups (p = 0.03 and p = 0.01, respectively). Data points show means and SEM, 95% confidence interval (n = 7 per group).
Fig 3.
Representative sections of liver parenchyma, stained with TUNEL.
(A) Representative section of tissue from a liver in the vehicle group. (B) Representative section of tissue from a liver in the F573 group. (C) Representative tissue from a liver in the anakinra and etanercept group. (D) Sample liver tissue from the BMX-001 group. Tissue was stained using fluorescein-12-dUTP dye.
Fig 4.
Reactive oxygen species quantification and caspase-3 activity.
A. Oxidative stress quantified by lipid peroxidation malondialdehyde (MDA) assay (p = 0.45) (B) Caspase-3 activity determined by comparison to the standard curve of pNA (p = 0.84), (C) Caspase-3 protein level quantification normalized to ß-actin from western blot (p = 0.10). Data columns show means and SEM, 95% confidence interval (n = 4, 6, and 4, respectively per group).
Fig 5.
(A) MPO staining calculated as percentage of total area per tissue section (p = 0.15). (B) F4/80 staining calculated as percentage of total area per tissue section (p = 0.047). Data columns show means and SEM, 95% confidence interval (n = 5 per group).
Fig 6.
Histological scoring of liver injury.
(A) All tissue slides were examined and scored by an independent expert pathologist for hemorrhage, necrosis, sinusoidal dilatation, and bile sequestration. Treated groups all demonstrated less injury, without reaching statistical significance (p = 0.11). Data columns show means and SEM, 95% confidence interval (n = 6 per group).
Fig 7.
Representative histologic sections of liver parenchyma, stained with hematoxylin and eosin.
(A) Representative section of tissue from a liver in the vehicle group. The curved black line indicates transition between normal tissue (left side of line) and necrotic liver (right of the line). (B) Representative section of tissue from a liver in the F573 group. (C) Representative tissue from the anakinra and etanercept group. (D) Liver tissue from the BMX-001 group. The curved black line indicates transition between normal tissue (left side of line) and necrotic liver (right of the line). All photographs are shown at 20X magnification. Arrows indicate areas of liver damage relevant to the scoring system, including necrosis and hemorrhage.
Fig 8.
Liver cytokine and biomarker analysis.
All treatment groups are compared to the vehicle group. (A) IFN-γ levels (Eta+Ana p = 0.006), (B) TNF-α levels (F573 p = 0.04); Eta+Ana p = 0.009), (C) IL-1 levels (Eta+Ana p = 0.47), (D) IL-6 levels (Eta+Ana p = 0.0003), (E) IL-10 levels (p = 0.09), (F) KC/GRO levels (Eta+Ana p = 0.0003), (G) IL-4 levels (Eta+Ana p = 0.04), (H) IL-5 levels (F573 p = 0.02). Data columns show means and SEM, 95% confidence interval (n = 6–8 per group).