Effect of diet and genotype on the lipidome of mice with altered lipoprotein metabolism

Figures

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  Figure 1. Effect of dietary treatment on mouse phenotype and global lipid pattern Body weight (A) and liver weight (B) at the end of the dietary treatments are shown. Histological measurement of atherosclerosis development (C) and representative hematoxylin and eosin staining of atherosclerotic plaques in the aortic sinus of mice fed CD and WD (D) are also shown, together with representative images of Oil Red O staining of WD-fed mouse livers (E). Scatterplots of the first two principal components (PCs) are shown for each sample, calculated from each analyte concentration in plasma (F), liver (G), and aorta (H). Bar length = 500 μm. Data are shown as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Significant differences within the same genotype (diet comparison) were determined by T-test, whereas differences among groups fed the same diet (genotype comparison) were determined by ANOVA followed by Tukey’s post-hoc test. n = 4–6 per group.

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  Figure 2. Lipid class concentrations in the three districts analyzed Concentration of lipid classes in plasma (A), liver (B), and aorta (C) are shown as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Significant differences within the same genotype (diet comparison) were determined by T-test, whereas differences among groups fed the same diet (genotype comparison) were determined by ANOVA followed by Tukey’s post-hoc test. n = 4–6 per group.

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  Figure 3. Overall effect of the Western diet on the lipid pathways analyzed The pathways involving the lipid classes measured in plasma, liver, and aorta of the three genotypes are represented. Triangles identify those lipid classes whose concentration was increased by WD in all genotypes.

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  Figure 4. Median changes of the 28 lipid species whose concentration is increased or decreased/unchanged by Western diet in all districts and genotypes Lipid species increased by WD vs. CD in plasma, liver and aorta of WT, Pcsk9-KO, and Ldlr-KO mice are shown with red bars, whereas those decreased by WD are shown with blue bars. Significant differences were determined by T test. ∗p < 0.05; ∗∗p < 0.01, ∗∗∗p < 0.001. n = 3–6 per group.

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  Figure 5. Overall effect of Pcsk9 deletion on the lipid pathways analyzed The pathways involving the lipid classes measured in plasma, liver, and aorta of Pcsk9-KO and WT mice are represented. Squares/diamonds identify decreases and triangles identify increases in those lipid classes whose concentration was significantly different between Pcsk9-KO and WT mice. Filled symbols refer to differences between CD-fed mice, chequered symbols highlight differences between WD-fed mice.

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  Figure 6. Plasma levels of the 32 lipid species whose concentration was lower in Pcsk9-KO vs. WT mice with both dietary treatments Significant differences were determined by T test. ∗p < 0.05; ∗∗p < 0.01, ∗∗∗p < 0.001. n = 3–6 per group.

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  Figure 7. Overall effect of Ldlr deletion on the lipid pathways analyzed The pathways involving the lipid classes measured in plasma, liver, and aorta of Ldlr-KO, Pcsk9-KO, and WT mice are represented. Larger circles identify those lipid classes whose concentration was significantly increased in Ldlr-KO mice vs. WT and Pcsk9-KO mice. Filled circles refer to differences among CD-fed mice, chequered circles highlight differences among WD-fed mice.

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  Figure 8. Median changes of the lipid species showing the highest increase in the aorta of Ldlr-KO mice vs. both WT and Pcsk9-KO mice on WD feeding Red bars indicate that the concentration is increased in the first comparison term, blue bars indicate that the concentration is reduced in the first comparison term. Significant differences were determined by T test. ∗p < 0.05; ∗∗p < 0.01, ∗∗∗p < 0.001. n = 3–6 per group. See also Figure S10.

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  Figure 9. Lipid species increased in plasma, liver, and aorta of WD-fed Ldlr-KO mice compared with WT and Pcsk9-KO mice Median changes (A) and absolute concentrations in plasma (B), liver (C) and aorta (D) are shown. Sixteen lipid species emerged from this analysis. Red bars indicate that the concentration is increased in the first comparison term, and blue bars indicate that the concentration is reduced in the first comparison term. Significant differences were determined by T test. ∗p < 0.05; ∗∗p < 0.01, ∗∗∗p < 0.001. n = 4–6 per group.

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  Figure 10. Absolute concentrations of the lipid species increased in the aorta of WD-fed Ldlr-KO mice compared with WD-fed WT and Pcsk9-KO mice and with CD-fed Ldlr-KO mice Forty-two lipid species emerged from this analysis. Significant differences were determined by T test. ∗p < 0.05; ∗∗p < 0.01, ∗∗∗p < 0.001. n = 3–6 per group.