Ecklonia stolonifera Okamura Extract Suppresses Myocardial Infarction-Induced Left Ventricular Systolic Dysfunction by Inhibiting p300-HAT Activity

Figures

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  Figure 1. PE-induced hypertrophic responses were suppressed by ESE treatment in cardiomyocytes. Primary cultured cardiomyocytes were treated with 100, 300, or 1000 μg/mL ESE and stimulated with 30 μM PE. (A) Immunofluorescence staining was performed using anti-α-actinin antibodies and Alexa Fluor 555-labeled anti-mouse IgG. Scale bar: 20 μm. (B) Cardiomyocyte surface area measurements. All data are presented as the mean ± SEM of three individual experiments. (C,D) Total RNA was extracted from the cells, and quantitative RT-PCR was performed for (C) ANF, (D) BNP, and HPRT1. Data are presented as the mean ± SEM of three individual experiments. (E) Histone fractions isolated from the cells were subjected to Western blotting using anti-acetyl-histone H3K9 antibodies and anti-histone H3 antibodies. (F) Quantified levels of acetylated histone H3K9 and total histone H3. Data are presented as the mean ± SEM of three individual experiments.

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  Figure 2. The effect of ESE on p300-HAT activity in vitro. (A) Samples were subjected to Western blotting with anti-acetyl-histone H3K9 antibodies and anti-histone H3 antibodies. Shown are quantified levels of acetylated histone H3K9 and total histone H3. Data are presented as the mean ± SEM of three individual experiments. (B) Concentration–response curve obtained by plotting acetyl-histone H3K9/histone H3 vs. common logarithm (concentrations). The IC50 value of ESE was 505 μg/mL. Quantified values of ESE are presented as the mean ± SEM of three individual experiments.

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  Figure 3. MI-induced systolic dysfunction was improved by ESE treatment in vivo. Cardiac function was measured by echocardiography at 8 weeks after treatment. (A) Representative images of echocardiography. (B) MI-induced increase in LV mass index was significantly suppressed by 1 g/kg of ESE. Data are presented as the mean ± SEM. (C) MI-induced increase in HW/TL was significantly suppressed by 1 g/kg of ESE. (D) Representative photographs of PSR-stained sections of LV myocardium from sham and MI rats. Scale bar: 2 mm.

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  Figure 4. Cardiac hypertrophy was suppressed by ESE treatment in rats with MI. (A) Representative images of HE-stained sections of cardiomyocytes from sham and MI rats. Magnification: ×400. Scale bar: 20 μm. (B) Myocardial cell diameter was measured for at least 50 cells in each rat. Data are presented as the mean ± SEM. (C,D) Quantitative RT-PCR data for (C) ANF, (D) BNP, and HPRT1. Data are presented as the mean ± SEM.

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  Figure 5. Cardiac perivascular fibrosis was suppressed by ESE treatment in rats with MI. (A) Representative images of PSR-stained perivascular fibrosis area of LV myocardium from sham and MI rats. Magnification: ×400. Scale bar: 50 μm. (B) Areas of perivascular fibrosis in the left ventricle were measured for at least three intramyocardial coronary arteries with a lumen size > 50μm. Data are presented as the mean ± SEM. (C,D) Quantitative RT-PCR data for (C) α-SMA, (D) Col 1A1, and HPRT1. Data are presented as the mean ± SEM.

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  Figure 6. The acetylation of histone H3K9 was suppressed by ESE treatment in LV of rats with MI. (A) Histone fractions from the rat hearts were subjected to Western blotting to assess acetylated histone H3K9 and total histone H3 levels. (B) Quantified levels of acetylated histone H3K9 and total histone H3. Data are presented as the mean ± SEM.