Effect of squalene on antioxidative parameters from the three experimental models.

<p>ROS levels in lipoprotein fractions from control and squalene treated mice expressed as arbitrary fluorescence units after incubation of lipoprotein fractions during 24 h with 2′,7′-dichlorofluorescein diacetate A) 0.8 µg LDL or VLDL, and 60 ng HDL from WT mice; B) 0.8 µg LDL and 60 ng HDL from <i>Apoa1</i>-deficient mice; C) 1.5 µg LDL or VLDL, and 60 ng HDL from <i>Apoe</i>-deficient mice. Each pool was assayed in triplicate. Individual plasma malondialdehyde levels from WT (D), <i>Apoa1</i>-(E) and <i>Apoe</i>-deficient mice (F). HDL-PON1 levels from WT (G), <i>Apoa1</i>-(H) and <i>Apoe</i>-deficient mice (I) with their representative Western blots. Each pool was assayed in triplicate. Results are shown as means ± SD. *p<0.05, **p<0.01 according to corrected unpaired t Welch's test.</p

Effect of the experimental diets on somatic variables in male mice of the three experimental models.

<p>Results are shown as mean values with their standard deviations. Statistical analysis was carried out using Mann Whitney U test.</p><p>* p<0.05 <i>vs</i> control.</p

Effect of dietary squalene supplementation on plasma parameters in male <i>Apoe</i>-deficient mice.

<p>Results are shown as mean values with their standard deviations. Statistical analysis was carried out using Mann Whitney U test.</p><p>*p<0.05,</p><p>**p<0.01 mean values were significantly different from those of the control diet, and</p>∧<p>p<0.05,</p>∧∧<p>p<0.01 mean values were significantly different between 0.25 and 1 g/kg squalene doses.</p

Relationship between hepatic gene expression and plasma cholesterol in wild type mice.

<p>Direct correlations among <i>Pcyox1</i> and <i>Pon2</i> gene expression (A), or plasma cholesterol (B), and <i>Lcat</i> and <i>Pon1</i> gene expression (C). Association analyses were carried out using Spearman's test for non- parametric distributions.</p

Effect of squalene on lipoproteins from the three experimental models.

<p>Plasma was obtained following 11 weeks of consuming control or squalene- enriched semipurified diets and after a four-hour fast. Two independent pools of all mice per experimental group were prepared, except for <i>Apoe</i>-deficient mice, where three plasma pools were utilized. Lipoproteins were separated by FPLC, and collected fractions analyzed for total cholesterol (A, G, L), esterified cholesterol (B, H, M), phosphatidylcholine (C, I, N), sphingomyelin (D, J, O), APOA1 (E, P) and APOA4 (F, K, Q). Representative profiles are shown from WT mice, left panels (control and squalene pools consisting of plasma from 6 and 7 mice, respectively), <i>Apoa1</i>-deficient mice, middle panels (n = 7 for control and n = 7 for squalene plasma pool) and Apoe-deficient mice, right panels (n = 13 for control, n = 13 for 0.25 g/kg and n = 14 for 1 g/kg squalene plasma pool). Fraction numbers 1–6 corresponded to VLDL/chylomicron remnants, 7–13 to low density lipoproteins, 14–21 to cholesterol-rich HDL and 22–27 to cholesterol-poor HDL (pHDL).</p

Changes in gene expression in small intestine fragments following gavage of 5 ml of virgin olive oil.

<p>Values expressed as means ± standard deviations. Data represent arbitrary units obtained with the RT-qPCR normalized to <i>Tbp</i>, <i>Ubc</i> and <i>Hprt</i> expressions for duodenum, jejunum and ileum, respectively. Statistical analysis was done using non-parametric one-way ANOVA according to Kruskal-Wallis test and unpaired Mann-Whitney U-test as post-hoc test. Superscripts (^a vs Control, ^b vs 4 h) indicate statistically significant differences (P<0.05).</p

Lipid composition of rat HDL following the consumption of 5-ml bolus of virgin olive oil.

<p>Values are expressed as means ± standard deviations of 2 pools per group. Statistical analysis was done using unpaired Mann-Whitney U test as post-hoc test. Superscripts (^a vs Control) indicate statistically significant differences (P<0.05).</p

Changes in hepatic steatosis in rats after gavage administration of fat in the form of virgin olive oil.

<p>Representative micrographs (×400 magnification) of livers from fasted animals (A) and 4 hours (B) and 8 hours (C) after receiving 5 ml of olive oil as a bolus. Liver sections (4 µm) were stained with hematoxylin and eosin and evaluated blindly. Morphometric changes in hepatic fat content (D) and analysis of triglyceride (E) and cholesterol (F) contents in rats where data are expressed as means ± SD for each group. Statistical analysis to evaluate dietary response was done using one-way ANOVA and the Mann-Whitney U test as post hoc test. ^a, P<0.05 vs control and ^b P<0.05 vs 4 h.</p

Changes in postprandial lipoproteins in rats after gavage administration of fat in the form of virgin olive oil.

<p>After a 16-h fast, male rats were subjected to oral gavage with virgin olive oil (16 ml kg⁻¹). Plasma lipoproteins were separated by FPLC and collected fractions analyzed for APOA1 (A), APOA4 (B), total cholesterol (C), esterified cholesterol (D), phosphatidylcholine (E) and sphingomyelin (F). A representative profile of one animal is shown. Fraction numbers 1–6 corresponded to VLDL/chylomicron remnants, 7–13 to low density lipoproteins, 14–18 to cholesterol-rich HDL and 19–24 to cholesterol-poor HDL.</p

Effect of CP-346086 or tyloxapol administration to rats on postprandial lipoprotein profile at eight hours.

<p>Rats were fasted overnight and administered vehicle, CP-346086 or tyloxapol. One hour later, they received a gavage dose of virgin olive oil and were sacrificed 8 hours after feeding. A representative profile of lipoprotein distribution according to APOA1 (A), APOA4 (B), total cholesterol (C), esterified cholesterol (D), free cholesterol (E), phosphatidylcholine (F), sphingomyelin (G) and triglyceride (H) contents.</p