Effects of ω-3 PUFA-Rich Oil Supplementation on Cardiovascular Morphology and Aortic Vascular Reactivity of Adult Male Rats Submitted to an Hypercholesterolemic Diet

Figures

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  Figure 1. Experimental design of induction of dyslipidemia and treatment with ω-3 PUFA-rich oil.

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  Figure 2. Total ion chromatogram (TIC) of methyl esters derived from ω-3 polyunsaturated fatty acids. Legend: x-axis: ‘Retention time (min)’; y-axis: ‘Relative intensity (%)’.

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  Figure 3. Photomicrographs of myocardial tissue from the CN56 group (A), CH56 group (B,C), and PUFA56 group (D–F). Hematoxylin–eosin (HE) stain (scale bar: 20 µm and 60 µm). (A): Rounded cross-sections of cardiac muscle cells with a centrally located nucleus. (B): Longitudinal section of myofiber disarray, with diffuse myodegeneration and hemorrhagic areas (*). (C): Detail of B showing foam cell (arrow). (D): Myofibers with a branched appearance, some showing vacuolar degeneration (circle) and infiltration of mononuclear cells. (E): Numerous mononuclear cells and fibroblasts, capillaries (arrows), and vacuolar degeneration areas (circle). (F): Detail of E showing foam cell (arrow) and fibroblasts (circle).

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  Figure 4. Photomicrographs of aorta from CN56 group (A,B), CH56 group (C,D), and PUFA56 group (E,F). Hematoxylin–eosin (HE) stain (scale bar: 100 µm and 50 µm). (A): This section shows the tunica media, composed of alternating layers of circumferentially arranged smooth muscle cells (arrow). (B): Detail of (A) showing wavy sheets of elastic fibers (*) and smooth muscle cells (arrow). (C): This section shows the disarrangement of the elastic fibers and the vacuolization of smooth muscle cells (circles) of the middle layer of the aorta. (D): Detail of (C) showing extracellular lipid (*), smooth muscle cell proliferation (black arrows), foam cell (arrowhead), and epithelial denudation (red arrows). (E,F): Note the reorganization of the histological structure of the artery, with more evident elastic fibers and a decrease in the vacuolar aspect of the smooth muscle cells (arrows).

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  Figure 5. Concentration–response curves induced by phenylephrine (A,D), acetylcholine (B,E) and sodium nitroprusside (C,F) in aortic artery rings from hypercholesterolemic rats treated with ω-3 PUFA-rich oil for 56 days, and Emax values thereof. Results are expressed as mean ± SEM (n = 6–10). Unpaired Student’s t-test. * p < 0.05, ** p < 0.01 and **** p < 0.0001 vs. CN56; #### p < 0.0001 vs. CH56.

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  Figure 6. Concentration–response curves induced by acetylcholine (10−9–10−5 M) in the absence or presence of pyrogallol (30 µM) in aortic artery rings from hypercholesterolemic rats treated with ω-3 PUFA-rich oil for 56 days. Results are expressed as mean ± SEM. Two-way ANOVA followed by Bonferroni posttest. ** p < 0.01 vs. CN56; **** p < 0.0001 vs. CH56. (A) CN56; (B) CH56; (C) PUFA56; (D) concentration–response curves for acetylcholine in the presence of pyrogallol.

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  Figure 7. Effects of ω-3 PUFA-rich oil on myeloperoxidase activity (A), nitrite concentration (B), superoxide dismutase activity (C), and catalase activity (D) in rat aortic tissue homogenates from hypercholesterolemic rats after supplementation for 56 days. Mean ± SEM (standard error of the mean) of 8 animals/group. Unpaired Student’s t-test. * p < 0.05 vs. CN56; # p < 0,05 vs. CH56.