Table 1.
Phytosterol and Alpha-tocopherol Levels in emulsions.
Table 2.
Emulsion USP <729> particle size analysis.
Fig 1.
Growth parameters between groups administered the PN diet and administration of fat emulsions formulated in the laboratory.
Fig 1A: Body masses were monitored every other day over the course of the enteral PN regimen. There were no differences in growth curves on linear regression analysis. Liver (Fig 1B), spleen (Fig 1C), and right kidney (Fig 1D) masses at euthanasia after 19 days of the PN diet. * = p<0.05 compared to PN FO group and PN FO+P group. N = 10 mice per treatment group. Statistical analysis of 1B, 1C, and 1D was performed with one-way ANOVA.
Fig 2.
Serum fatty acid profiles after 19 days of the PN diet with administration of fat emulsions formulated in the laboratory.
Fig 2A: Triene to tetraene ratios. Only mice receiving no fat source (PN+saline) met biochemical criteria for EFAD. Distribution of the main omega-6 (Fig 2B) and omega-3 fatty acids (Fig 2C) reflect the fat source administered to each group. N = 3 samples per treatment group randomly selected. Statistical analysis was performed with one-way ANOVA. * = p<0.05 with respect to the PN FO group.
Fig 3.
Normalization of hepatic architecture and hepatic fat accumulation with addition of α-tocopherol to SO in an intravenous fat source with the enteral PN diet.
Representative Hematoxylin and Eosin (H&E, Fig 3A) and Oil Red O (Fig 3B) images demonstrating hepatic architecture and hepatic fat accumulation, respectively. N = 10 samples per treatment group for H&E (Fig 3A), and n = 3 representative samples for Oil Red O (Fig 3B). Images are 100X magnification (10X objective, 10X eyepiece).
Fig 4.
Hepatic gene expression of ACC2 and PPARg are dysregulated in the presence of enteral PN-induced liver injury.
Expression of ACC2 (Fig 4A) and PPARγ (Fig 4B) is dysregulated by the fat-free PN diet and the PN diet with IV SO as a fat source, and normalized by IV FO and by the addition of α-tocopherol to SO as fat sources. Gene expression is measured as fold-difference compared to the chow-fed group. N = 5 samples per treatment group, each performed in technical duplicate. Statistical analysis was performed with one-way ANOVA.
Fig 5.
Hepatic protein expression of ACC2 and PPARg are dysregulated in the presence of enteral PN-induced liver injury.
Protein levels of ACC2 (Fig 5A) and PPARγ (Fig 5B) are upregulated by the fat-free PN diet and the PN diet with IV SO as a fat source, and normalized by IV FO and by the addition of α-tocopherol to SO as fat sources. Protein levels were quantified comparatively by normalizing each group to the corresponding beta-actin level and comparing to the chow-fed group. Western Blots performed in biological duplicate for each group. Statistical analysis was performed with one-way ANOVA. Fig 5C: Western Blot image.
Fig 6.
Expression of regulators of bile homeostasis with expression altered by phytosterol-containing intravenous lipids, and at least partially normalized by α-tocopherol.
Data are expressed as fold difference in expression compared to the chow-fed control group. A) Cyp7a1 = regulator of bile acid synthesis; B) FXR = farsenoid X receptor; C) SHP = small heterodimer protein. N = 5 samples per group each performed in technical duplicate, and analysis was performed with single-factor ANOVA.