Figure 1.
Oleic acid increase is highest in MAT.
Mass spectrometry analysis of (A) fatty-acid composition (expressed as a percentage of total fatty acid) in the EAT and MAT following three control or 60% HMF feeding. (B) Change in the percentage of palmitic (PA) and oleic (OA) acid out of total fatty-acids expressed as the difference between control and HMF-fed mice. (N = 5–6).
Table 1.
Fatty acid composition of milk-fat.
Table 2.
Fatty acid composition in adipose tissues following short term control or HMF feeding.
Figure 2.
Oleic acid induces M2 macrophage markers in
Arginase-1 (Arg1) and TNFα gene expression fold change from (A) RAW 264.7 macrophages or (B) bone marrow derived macrophages (BMDM) cultured overnight with oleic (OA) or palmitic (PA) acid (500 µM). (N = 3,4) (C) CD206, MGL1 and KLF4 gene expression fold change in RAW 264.7 macrophages following overnight culture in OA (500 µM). (D) Arg1 and CD206 gene expression fold change from RAW 264.7 macrophages cultured in OA (200 µM) and PA (as labeled). Control cells were cultured in BSA and ethanol. (N = 3).
Figure 3.
Dietary oleic acid increases M2 macrophage markers in the MAT.
(A) Change in body-weight during a three day, twice daily, oral gavage with control or purified OA. (B) F4/80 gene expression (qPCR relative to GAPDH) in mesenteric adipose tissue (MAT) or epididymal adipose tissue (EAT) from mice given control or OA gavage. (C) Gene expression (qPCR relative to GAPDH) analysis of M2 and M1 macrophage markers in MAT from gavaged mice. (N = 10, 12).
Figure 4.
Metabolic and body composition changes in response to 3 day HMF feeding.
The mouse metabolic research unit was utilized to analyze (A) body-weight changes, (B) respiratory exchange ratio (RER) and (C&D) food-intake in mice fed a three day control or 42% HMF diet. (N = 5) (E) Fat and lean mass were measured using MRI. (N = 5).
Figure 5.
Diet-induced macrophage changes in MAT and EAT.
(A) F4/80 expression (qPCR relative to GAPDH) in MAT and EAT (n = 4–8). (B) MCP-1 expression (qPCR relative to GAPDH) in MAT and EAT (n = 4–8). (C) Changes in MCP-1 protein detected by ELISA in extracts of adipose tissues and expressed as ratios of HMF diet over control diet adipose tissue levels in EAT and MAT. (n = 4–8) (D) Typical flow cytometry pattern of the SVF of the MAT showing (a) F4/80Int CD206+ and (b) F4/80HI CD206− macrophage populations. (E) Changes in the F4/80Int CD206+ macrophage population expressed as ratios of HMF diet over control diet adipose tissue cells/gm in EAT and MAT (n = 6). (F) Representative flow cytometry of the F4/80HI CD206− macrophage population in MAT showing CD11b expression with control and HMF diets; and graph of the percentage of these cells in the total macrophage population within the stromavascular fraction of MAT from control and HMF diets. (G) Changes in the F4/80HI CD206− CD11bHi macrophages expressed as ratios of cells/g in HMF diet over control diet EAT and MAT.
Figure 6.
HMF diet induces peritoneal cell adhesion.
(A) F4/80 and CD206 expression, by flow cytometry, on peritoneal macrophages, and SVF macrophages isolated from unwashed or washed MAT. F4/80Int CD206+ (a) and F4/80HI CD206− (b) macrophage populations are boxed. (B) Total cell counts (per mouse) of peritoneal cells (PCs) or macrophages (Macs) isolated by peritoneal lavage following three day control or milk-fat feeding. (N = 3) (C) C57BL6, MCP-1−/−, or ICAM-1−/− mice were fed control or HMF diet. F4/80 gene expression from HMF unwashed MAT is expressed as fold change relative to mice on control diet. (N = 5–8) (D) F4/80 and MCP-1 gene expression (qPCR, relative to Gapdh) in the washed MAT of mice fed a three day control or HMF diet. (N = 10,11).
Figure 7.
Dietary oleic acid increases M2 markers in the washed MAT.
Gene expression analysis for (A) F4/80 and MGL1, (B) subsets of M2 macrophage markers (qPCR, relative to Gapdh), in washed MAT from mice given control gavage or oleic acid gavage twice daily for three days. (N = 10, 12).