Blockade of Inflammatory Markers Attenuates Cardiac Remodeling and Fibrosis in Rats with Supravalvular Aortic Stenosis

Figures

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  Figure 1. Experimental design. Wistar rats were submitted to an aortic stenosis (AS) protocol or sham surgery, and 21 weeks after surgery, all rats were submitted to a physical capacity test and transthoracic echocardiography for division of the groups. After 24 weeks of surgery, animals were treated with DEX (50 μg/kg/day, s.c.), ramipril (10 mg/kg/day, gavage) or saline for 14 days. After the experimental protocol, the animals were submitted to an echocardiographic examination and euthanasia.

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  Figure 2. Black line (A) and black bar (B) represent the sham group, while gray line (A) and gray bar (B) represent aortic stenosis rats. Analysis of the survival rate (A) measured in both groups sham (n = 28) and aortic stenosis (AS, n = 113) and results of maximal physical capacity test (B) measured in both groups sham (n = 20) and aortic stenosis (AS, n = 23). Results are presented as mean ± standard error of the mean, significance: * p < 0.05, ** p < 0.01 vs. sham group.

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  Figure 3. Upper panel: Representative images of hematoxylin and eosin (H and E, (A,B)) and Picrosirius Red (C) staining for each group; (A) longitudinal section showing the myocytes as demonstrated by the black line; (B) transversal section showing the capillaries as indicated by the black arrow; and (C) transversal sections showing the collagen deposition, in which collagen fibers can be seen in red and cardiac musculature in yellow. Bar: 50 µm, increase of 400× (A,B) and 100 µm, increase of 200× (C). Bottom panel: From hematoxylin–eosin technique, analyses of •myocyte diameter (µm, (D)) and •capillary density on the left ventricle (n/mm2, (E)). From picrosirius red staining images, densitometric results of left ventricle collagen deposition area (μm2, (F)) and % of collagen area (G) in all analyzed groups: sham (S, n = 12) and aortic stenosis (AS, n = 9). Results are presented as mean ± standard error of the mean. • means parametric test. Significance: * p < 0.05, vs. sham group.

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  Figure 4. Analysis of cardiac inflammatory markers: interleukin 1α (IL1-α), interleukin 1 β (•IL1-β), interleukin 4 (•IL-4), interleukin 6 (IL-6), interleukin 10 (•IL-10), interleukin 12 (IL-12), interferon gama (IFN-γ), transforming growth factor β (TGF-β), tumor necrosis factor α (•TNF-α) and vascular endothelial growth factor (•VEGF). Sham (S, n = 10) and aortic stenosis (AS, n = 9). Results are presented as mean ± standard error of the mean. • means parametric test. Significance: * p < 0.05, vs. sham group.

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  Figure 5. Analysis of cardiac inflammatory markers: interleukin 1α (IL1-α), interleukin 1 β (IL1-β), interleukin 4 (•IL-4), interleukin 6 (•IL-6), interleukin 10 (IL-10), interleukin 12 (IL-12), interferon gama (•IFN-γ), transforming growth factor β (TGFβ), tumor necrosis factor α (•TNFα) and vascular endothelial growth factor (•VEGF). Aortic stenosis (AS, n = 9), aortic stenosis treated with DEX (ASD, n = 9) and aortic stenosis treated with ramipril (ASR, n = 9). Results are presented as mean ± standard error of the mean. • means parametric test. Significance: + p < 0.05 vs. AS group.

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  Figure 6. Upper panel: Representative images of hematoxylin and eosin (H and E, (A,B)) and Picrosirius Red (C) staining for each group; (A) longitudinal section showing the myocytes as demonstrated by the black line; (B) transversal section showing the capillaries as indicated by the black arrow; and (C) transversal sections showing the collagen deposition, in which collagen fibers can be seen in red and cardiac musculature in yellow. Bar: 50 µm/increase of 400× (A,B) and 100 µm/ increase of 200× (C). Bottom panel: From hematoxylin–eosin technique, analyses of •myocyte diameter (µm, (D)) and capillary density on the left ventricle (n/mm2, (E)). From picrosirius red staining images, densitometric results of left ventricle collagen deposition area (μm2, (F)) and % of collagen area (G) in all analyzed groups: aortic stenosis (AS, n = 9), aortic stenosis treated with DEX (ASD, n = 9) and aortic stenosis treated with ramipril (ASR, n = 9). Results are presented as mean ± standard error of the mean. • means parametric test. Significance: + p < 0.05 vs. AS group.