The Effects of Diabetes Induction on the Rat Heart: Differences in Oxidative Stress, Inflammatory Cells, and Fibrosis between Subendocardial and Interstitial Myocardial Areas

Figures

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  Figure 1. Glycemia (a) and insulin (b) serum concentrations and RAGE protein expression (c) after 8 weeks of diabetes induction. ∗p < 0.05 and †p < 0.001 versus the CT group. Data expressed in mean ± SEM in all plots.

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  Figure 2. (a) Quantitative analysis of microfluorotopography of DHE oxidation products performed 8 weeks after diabetes induction. ∗p < 0.01 versus the CT SEN area. Data expressed in mean ± SEM. (b) Representative photomicrograph of SEN and INT areas, showing microfluorotopography of DHE oxidation products. Red staining indicates the fluorescence signal by DHE under 400x magnification. Bars: 50 μm. Western blot protein expression analysis depicting antioxidant enzymes catalase (c) and SOD1 (d) performed 8 weeks after diabetes induction. †p < 0.001 versus the CT group. Data expressed in mean ± SEM.

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  Figure 3. (a) Total inflammatory cell count in SEN and INT areas. HE-stained sections, under 400x magnification. θp < 0.05 versus the DM SEN area; ∗p < 0.01 versus the CT SEN area, #p < 0.01 versus the DM SEN area; †p < 0.01 versus the CT INT area. Data expressed in mean ± SEM. Western blot protein expression analysis depicting CD68 (b), CD3 (c), MCP-1 (d), TNF-α (e), IL1-β (f), and IL-6 (g) was performed 8 weeks after diabetes induction. ∗p < 0.05 and †p < 0.001 versus the CT group. Data expressed in mean ± SEM in all plots.

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  Figure 4. (a) Representative photomicrograph of myocardial necrosis in SEN and INT areas. HE-stained sections under 400x magnification. Arrows indicate nuclear pyknosis, karyolysis, cytoplasmatic changes, and hypereosinophilia. Bars: 50 μm. Western blot protein expression analysis depicting proapoptotic caspase 3 (b) and BAX (c) and antiapoptotic Bcl-2 (d) and HIF1-α (e) performed 8 weeks after diabetes induction. #p < 0.01 and †p < 0.001 versus the CT group. Data expressed in mean ± SEM.

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  Figure 5. (a) Quantitative analysis of collagen volume fraction in SEN and INT areas, performed 8 weeks after diabetes induction. θp < 0.05 versus the DM SEN area; ∗p < 0.01 versus the CT SEN area; †p < 0.01 versus the CT INT area. Data expressed in mean ± SEM. (b) Representative photomicrograph of SEN and INT areas, showing myocardial fibrosis in Masson's trichrome stain. Blue staining indicates the myocardial fibrosis under 200x magnification. Bars: 100 μm. Western blot protein expression analysis depicting collagen I (c) and collagen III (d), performed 8 weeks after diabetes induction. ∗p < 0.05 versus the CT group. Data expressed in mean ± SEM in all plots.

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  Figure 6. Pearson's correlation between ROS generation (a, b), inflammatory cells (c, d), myocardial fibrosis (e, f) in SEN and INT areas, respectively, versus E/A wave, performed 8 weeks after diabetes induction.