Pomegranate Extract Enhances Endothelium-Dependent Coronary Relaxation in Isolated Perfused Hearts from Spontaneously Hypertensive Ovariectomized Rats

Figures

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  FIGURE 1. Systolic blood pressure (SBP) at the beginning (0 day) and end of treatment (30 days). Vehicle (V) and Pomegranate hydroalcoholic extract (PHE) in Sham group (A) and Ovariectomized group (OVX) (B). Values are expressed as mean ± SEM; n = 10–16 animals per group. ∗p < 0.05 compared to 0 Day values, #p < 0.05 compared to the Vehicle group.

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  FIGURE 2. Baseline coronary perfusion pressure (CPP) after the treatment. Sham treated with Vehicle (Sham-V), Sham treated with Pomegranate hydroalcoholic extract (Sham-PHE), Ovariectomized treated with Vehicle (OVX-V) and Ovariectomized treated with Pomegranate hydroalcoholic extract (OVX-PHE) groups. Values are expressed as mean ± SEM; n = 9–12 animals per group. ∗∗p < 0.01 compared to the Sham-V group.

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  FIGURE 3. Dose-response curves to bradykinin (0,1–1000 ng) in isolated hearts. Sham group (A) and OVX group (B) before (closed symbols) and after (open symbols) perfusion with L-NAME (100 μM) for 20 min. Data were expressed as mean ± SEM; n = 6–10 animals per group. ∗p < 0.05 compared with the vehicle group before L-NAME. #p < 0.05 compared with the both groups after L-NAME.

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  FIGURE 4. Analysis of protein expression in Sham and OVX rats. (A) p-eNOS (Ser1177), (B) p-NOS (Thr495) and (C) total eNOS in Sham-V, Sham-PHE, OVX-V and OVX-PHE. Each sample was obtained from 1 vessel from coronary septal arteries, and the analyses of different proteins were made in the same gels using isolated strip buffer. However, the organization of the groups not allowed us to remove a representative image of the blots in a single gel. Thus, for better clarification we reorganized the blots separately. Data were expressed as mean ± SEM; n = 5–10 per group. ∗p < 0.05 compared to Sham-V group.

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  FIGURE 5. Analysis of protein expression in Sham and OVX rats. (A) p-AKT (Ser 473), (B) total AKT in Sham-V, Sham-PHE, OVX-V, and OVX-PHE. Each sample was obtained from 1 vessel from coronary septal arteries, and the analyses of different proteins were made in the same gels using isolated strip buffer. However, the organization of the groups not allowed us to remove a representative image of the blots in a single gel. Thus, for better clarification we reorganized the blots separately. Data were expressed as mean ± SEM; n = 5–10 per group.

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  FIGURE 6. Analysis of protein expression in Sham and OVX rats. (A) superoxide dismutase, SOD-2 and (B) catalase in Sham-V, Sham-PHE, OVX-V, and OVX-PHE. Data were expressed as mean ± SEM; n = 4–7 per group. #p < 0.05 compared with OVX-V group.

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  FIGURE 7. Influence of pomegranate peel extract treatment on superoxide anion production in coronary arteries. The top panels (A–D) show higher fluorescence intensity (red) in OVX-V rats when compared both Sham and OVX-PHE rats using dihydroethidium (DHE) staining. Left panel, vehicle treatment and right panel, pomegranate hydroalcoholic extract treatment (PHE). (E) The bar graph shows the average DHE fluorescence (AU: arbitrary units) comparing all groups (n = 9–12). Values are means ± SEM. ∗∗p < 0.01 compared with Sham, and ##p < 0.01 compared with OVX-PHE group. Scale bar: 50 μm.

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  FIGURE 8. Analysis of plasma nitrite levels. Values in Sham-V, Sham-PHE, OVX-V and OVX-PHE. Data were expressed as mean ± SEM; n = 6–12 animals per group. ∗p < 0.05 compared to the Sham-V group. #p < 0.05 compared with OVX-V group.