Dietary Effects on Monocyte Phenotypes in Subjects With Hypertriglyceridemia and Metabolic Syndrome

Figures

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  Figure 1. Study Design In this randomized crossover study, 19 patients with hypertriglyceridemia and metabolic syndrome received a high–saturated fat diet and a low–saturated fat diet in a randomized order separated by a 4- to 6-week washout period. Each diet period included 2 study phases: a 4-day diet phase (days 1-4) and a postprandial phase (day 5). Blood was drawn on day 1 after an overnight fast (before diet intervention) and on day 5 fasting (0 hour) and 4 and 6 hours after a test meal, which contained ∼900 kcal and was similar in composition to the respective study diet. White arrows indicate fasting blood draws; closed arrows indicate postprandial blood draws; dashed lines indicate washout period; and the solid line indicates subsequent analyses.

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  Figure 2. Biochemical Profiles (A) Plasma levels of triglyceride and low-density lipoprotein triglyceride (LDL-TG). (B) Plasma levels of total cholesterol; LDL cholesterol (LDL-C), including small dense LDL-C (sdLDL-C) and large buoyant LDL-C (lbLDL-C); remnant-like particle cholesterol (RLP-C); and high-density lipoprotein cholesterol (HDL-C). (C) Plasma levels of apolipoprotein CIII (apoCIII). (D) Plasma levels of high-sensitivity C-reactive protein (hsCRP) and glycoprotein acetylation (GlycA). (E) Plasma levels of glucose, insulin, and homeostatic model assessment for insulin resistance (HOMA-IR). (F) Plasma levels of total fatty acids and percentages of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) in total fatty acids. Values are mean ± SEM. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 for main effects; #P < 0.05, ##P < 0.01, ###P < 0.001 post hoc test for pairwise comparisons between diets at indicated time points (day 1 [D1] fasting; day 5 [D5] fasting [0] and 4 hours [4] and 6 hours [6] after a test meal). HSFD = high–saturated fat diet; LSFD = low–saturated fat diet.

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  Figure 3. Changes in Monocyte Counts, Subset Frequencies, and Lipid Accumulation (A) Total monocyte counts. (B) Representative flow cytometric examples and frequencies of monocyte subsets: CD14++/CD16– classical (cM), CD14+/CD16+ intermediate (iM), and CD14dim/CD16+ nonclassical (nM) monocytes. (C) Side scatter (SSC), representing cell granularity/lipid droplets, and Nile Red staining for lipids (indicated by relative mean fluorescence intensity [MFI] of Nile Red staining) of monocyte subsets. Values are mean ± SEM. ∗P ≤ 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 for main effects; #P < 0.05 post hoc test for pairwise comparisons between diets at indicated time points. D5 = day 5; FSC = forward scatter; other abbreviations as in Figure 2.

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  Figure 4. Adhesion Molecules and Adhesion Capacity of Monocytes Expression levels, indicated by MFI, of adhesion molecules, including CD11c (A), CD11a (B), and very late activation antigen-4 (VLA4) (C), on monocyte subsets examined by flow cytometry. (D) Firm adhesion of monocyte subsets on vascular cell adhesion molecule-1 examined by the ex vivo micro-flow adhesion assay. Values are mean ± SEM. ∗P ≤ 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 for main effects; #P < 0.05, ##P < 0.01 post hoc test for pairwise comparisons between diets at indicated time points. Abbreviations as in Figures 2 and 3.

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  Figure 5. Monocyte CD36 Levels and oxLDL Uptake Ex Vivo (A) Expression levels of CD36, indicated by MFI, on monocyte subsets examined by flow cytometry. (B) Oxidized low-density lipoprotein (oxLDL) uptake, presented as DiI (oxLDL) MFI, by monocyte subsets examined by using flow cytometry after coincubation of DiI-labeled oxLDL with monocytes ex vivo for 1 hour. Values are mean ± SEM. ∗P < 0.05, ∗∗∗P < 0.001 for main effects; #P < 0.05 post hoc test for pairwise comparisons between diets at indicated time points. Abbreviations as in Figures 2 and 3.

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  Figure 6. Correlations Between Monocyte Markers and Plasma Levels of Lipids (A) Data from all time points and both diets combined. (B) Data according to diet but combined from all time points for each diet. The x-axis indicates monocyte markers, and the y-axis indicates lipid variables. DHA = docosahexaenoic acid; FA = fatty acid; HDL-C = high-density lipoprotein cholesterol; LA = linoleic acid; TC = total cholesterol; TG = triglyceride; other abbreviations as in Figures 2 and 3.