Figure 1.
Histological liver representation of parenteral nutrition-associated liver disease patients.
Hematoxilin & eosin and masson's trichrome staining of liver biopsies taken for diagnosis purposes. The images are representative of patients with non-alcoholic liver hepatic disease score (NAS) ≤6 and patients with NAS>6 who initiated Omegaven® treatment.
Table 1.
Baseline clinical characteristics.
Table 2.
Dynamic laboratory values during home parenteral nutrition (PN).
Table 3.
Hepatic and inflammatory marker values in healthy subjects, and patients after four months of Omegaven® treatment, and patients with chronic ClinOleic®, Lipofundin® and SMOFlipid® based-home parenteral nutrition. *p<0.05 vs. values of healthy and Omegaven®.
Figure 2.
Effect of different commercial parenteral lipid emulsions on Lipopolysaccharide (LPS)-induced TGFβ1 and MMP-9 secretion in human monocytes.
Human monocytes were isolated from healthy subjects and incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by LPS 1 µg/mL stimulation for additional 24 hours. (A) TGFβ1 and (B) MMP-9 were measured in cell culture supernatants. The effect of lipid emulsions without stimulus was tested at 1/10 dilution. Results are expressed as means ± SEM of six independent experiments. *p<0.05 related to the control group. #p<0.05 values below stimulus; ⊥p<0.05 values above the stimulus.
Figure 3.
Effect of different commercial parenteral lipid emulsions on Lipopolysaccharide (LPS)-induced IL-6 and TNFα secretion in human monocytes.
Human monocytes were isolated from healthy subjects and incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by LPS 1 µg/mL stimulation for additional 24 hours. (A) IL-6 and (B) TNFα were measured in cell culture supernatants. The effect of lipid emulsions without stimulus was tested at 1/10 dilution. Results are expressed as means ± SEM of six independent experiments. *p<0.05 related to the control group. #p<0.05 values below stimulus; ⊥p<0.05 values above the stimulus.
Figure 4.
Effect of different commercial parenteral lipid emulsions on Lipopolysaccharide (LPS)-induced IL-8 and IL-1β secretion in human monocytes.
Human monocytes were isolated from healthy subjects and incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by LPS 1 µg/mL stimulation for additional 24 hours. (A) IL-8 and (B) IL-1β were measured in cell culture supernatants. The effect of lipid emulsions without stimulus was tested at 1/10 dilution. Results are expressed as means ± SEM of six independent experiments. One-way ANOVA was followed by the post hoc Bonferroni test. *p<0.05 related to the control group. #p<0.05 values below stimulus; ⊥p<0.05 values above the stimulus.
Figure 5.
Effect of different commercial parenteral lipid emulsions on Lipopolysaccharide (LPS)-induced GM-CSF and IL-5 secretion in human monocytes.
Human monocytes were isolated from healthy subjects and incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by LPS 1 µg/mL stimulation for additional 24 hours. (A) GM-CSF and (B) IL-5 were measured in cell culture supernatants. The effect of lipid emulsions without stimulus was tested at 1/10 dilution. Results are expressed as means ± SEM of six independent experiments. One-way ANOVA was followed by the post hoc Bonferroni test. *p<0.05 related to the control group. #p<0.05 values below stimulus; ⊥p<0.05 values above the stimulus.
Figure 6.
Effect of different commercial parenteral lipid emulsions on Lipopolysaccharide (LPS)-induced IL-4 and IL-10 secretion in human monocytes.
Human monocytes were isolated from healthy subjects and incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by LPS 1 µg/mL stimulation for additional 24 hours. (A) IL-4 and (B) IL-10 were measured in cell culture supernatants. The effect of lipid emulsions without stimulus was tested at 1/10 dilution. Results are expressed as means ± SEM of six independent experiments. One-way ANOVA was followed by the post hoc Bonferroni test. *p<0.05 related to the control group. #p<0.05 values below stimulus; ⊥p<0.05 values above the stimulus.
Figure 7.
Effect of different commercial parenteral lipid emulsions on Lipopolysaccharide (LPS)-induced IL-2 and IL-12 secretion in human monocytes.
Human monocytes were isolated from healthy subjects and incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by LPS 1 µg/mL stimulation for additional 24 hours. (A) IL-2 and (B) IL-12 were measured in cell culture supernatants. The effect of lipid emulsions without stimulus was tested at 1/10 dilution. Results are expressed as means ± SEM of six independent experiments. One-way ANOVA was followed by the post hoc Bonferroni test. *p<0.05 related to the control group. #p<0.05 values below stimulus; ⊥p<0.05 values above the stimulus.
Figure 8.
Effect of different lipid emulsions on the expression of myofibroblast markers.
Human liver epithelial cell line THLE-3 was incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at 1/100 dilutions for 72 hours. Representative visible morphology and immunofluorescences for alpha smooth muscle actin (αSMA) and collagen type I (col type I) are showed.
Figure 9.
Omegaven® inhibits myofibroblast markers induced by TGFβ1.
Human liver epithelial cell line THLE-3 was incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by TGFβ1 5 ng/mL stimulation for additional 72 hours. (A) Visible morphology and immunofluorescence for alpha smooth muscle actin (αSMA) and collagen type I (col type I) distribution and expression. B) Expression of mRNA of αSMA and col type I. Scale bar: 10 µm. Results are expressed as means ± SEM of six independent experiments. *p<0.05 related to the control group. #p<0.05 related to the stimulus.
Figure 10.
Omegaven® inhibits epithelial to mesenchymal transition induced by TGFβ1.
Human liver epithelial cell line THLE-3 was incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by TGFβ1 5 ng/mL stimulation for additional 72 hours (A and B) or 25 min (C). (A) Expression of mRNA of ZO-1 and (B) E-cadherin. (C) Phosphorylation of Samd3, ERK1/2 and Akt and nuclear expression of β-catenin. Representative western blot are showed and quantified in graphic bars. Results are expressed as means ± SEM of six independent experiments. *p<0.05 related to the control group. #p<0.05 related to the stimulus.
Figure 11.
Omegaven® inhibits transcription factor an markers related with epithelial to mesenchymal transition induced by TGFβ1.
Human liver epithelial cell line THLE-3 was incubated in presence or absence of lipid emulsions Omegaven® 10%, Lipofundin MCT/LCT® 20%, ClinOleic® 20% or SMOFlipid® 20% at different dilutions, for 30 min followed by TGFβ1 5 ng/mL stimulation for additional 72 hours. Expression of mRNA of Snail, Slug, and vimentin. Results are expressed as means ± SEM of six independent experiments. One-way ANOVA was followed by the post hoc Bonferroni test. *p<0.05 related to the control group. #p<0.05 related to the stimulus.