Effects of Three-Month Feeding High Fat Diets with Different Fatty Acid Composition on Myocardial Proteome in Mice

Figures

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  Figure 1. Representative light micrographs of H&E-stained (A–D) and Mallory’s trichrome-stained (E–H) ventricle mid-region heart cross sections after STD (A,E), SFA (B,F), 14:1 (C,G) and 5:1 (D,H) diets. Scale bar—50 µm. STD – standard diet; SFA – high fat diet rich in saturated fatty acids; 14:1 – high fat diet rich in polyunsaturated fatty acids (PUFA) with LA/ALA ratio ~14:1; 5:1 – high fat diet rich in (PUFA) with LA/ALA ratio 5:1.

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  Figure 2. Representative two-dimensional electrophoresis gel image of the mouse heart proteome. Differentially expressed proteins between the animals fed standard diet (STD) and experimental high fat diets are indicated by numbers. Protein spot numbers refer to those presented in the heat map (Figure 3) and Table 3.

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  Figure 3. Heat map representing the identified proteins according to the magnitude of fold-change in the myocardium of mice fed high-fat diets with different fatty acid composition (SFA, n6/n3 PUFA 14:1, n6/n3 PUFA 14:1) compared to group of mice fed a standard diet (STD). Proteins were grouped according to their involvement in biological processes. Spot numbers refer to those presented on the 2-D proteome map (Figure 2) and Table 3. TCA - tricarboxylic acid cycle.

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  Figure 4. The ClueGO gene network of significantly altered pathways in cardiac muscle after 3 months of high-fat diets ingestion. Each node represents the Gene Ontology (GO) biological process/pathways and colors represent the GO group.

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  Figure 5. Overview of the effects of high-fat diets on the cardiomyocyte proteome. Identified proteins are shown according to the magnitude of fold-change in comparison to the STD group; red indicates upregulated proteins and green downregulated proteins in the myocardium of mice. EETs—epoxyeicosatrienoic acids; DHETs—dihydroxyeicosatrienoic acids; EEQs—epoxyeicosatetraenoic acids; DHEQs—eicosatetraenoic acids; EDPs—epoxydocosapentaenoic acids; DiHDPAs—dihydroxydocosapentaenoic acids; AMPK—AMP-activated kinase.

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  Figure 6. Validation of proteomic data obtained using 2-DE combined with MALDI-ToF MS. Protein expression levels of (a) short-chain specific acyl-CoA dehydrogenase, mitochondrial—ACADS (b) bifunctional epoxide hydrolase 2—sEH (c) superoxide dismutase 1—SOD1 (d) malate dehydrogenase, mitochondrial—MDH2. Western blots represent the murine myocardium from the control (STD) and experimental groups: SFA, 14:1 and 5:1.