The Mechanism Underlying the Protective Effects of Tannic Acid Against Isoproterenol-Induced Myocardial Fibrosis in Mice

Figures

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  Figure 1. Tannic acid (TA) structure.

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  Figure 2. The effects of TA on serum levels of creatine kinase (CK) (A), lactate dehydrogenase (LDH) (B), and CK-MB (C). Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (*P < 0.05 and **P < 0.01).

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  Figure 3. The effects of tannic acid (TA) on mitochondrial enzyme activities. Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (*P < 0.05 and **P < 0.01).

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  Figure 4. The effects of tannic acid (TA) on cardiac fibrosis as surveyed by H&E staining. The staining is shown for the CONT (A), isoproterenol (ISO) (B), PRO (C), low-dose TA (L-TA) (D), and high-dose TA (H-TA) (E) groups (H&E, 400×; bar 50 μm). Pay attention note the cardiomyocytes’ derangement distribution (arrows) as revealed with H&E staining. (F) Quantification of the H&E staining expressed as the proportion of the area positive for collagen fibers. Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to ISO group (**P < 0.01).

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  Figure 5. The effects of tannic acid (TA) on cardiac fibrosis as surveyed by Sirius red staining. The staining is shown for the CONT (A), isoproterenol (ISO) (B), PRO (C), low-dose TA (L-TA) (D), and high-dose TA (H-TA) (E) groups (200×; bar 100 μm). (F) Quantification of Sirius red staining expressed as the proportion of the positive area for collagen fibers. Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to ISO group (*P < 0.05 and**P < 0.01).

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  Figure 6. The effects of tannic acid (TA) on myocardial fibrosis as surveyed by Masson's trichrome staining. The staining is shown for the CONT (A), isoproterenol (ISO) (B), PRO (C), low-dose TA (L-TA) (D), and high-dose TA (H-TA) (E) groups (Masson's trichrome staining, 400×; bar 50 μm). (F) Quantification of Masson's trichrome staining expressed as the proportion of the positive area for collagen fibers. Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to the ISO group (**P < 0.01).

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  Figure 7. The effects of tannic acid (TA) treatment on Bax, Bcl-2, and basic fibroblast growth factor (bFGF) expression in heart observed after immunohistochemical dispose of. The morphological locations and percent areas of Bax (A1-6), Bcl-2 (B1-6), and bFGF (C1-6) expressions are shown (immunohistochemical, 400×; bar 50 μm). Positive expressions of Bax, Bcl-2, and bFGF are indicated by arrows. Data represent the mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (*P < 0.05 and **P < 0.01).

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  Figure 8. The effects of tannic acid (TA) treatment on Bax, Bcl-2, and caspase-3 expressions as shown by Western blotting (WB) analysis. The band intensities of Bax/Bcl-2 (A, B) and caspase-3 (C, D) were analyzed by normalization to β-actin. Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (*P < 0.05 and **P < 0.01).

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  Figure 9. The effects of tannic acid (TA) treatment on atrial natriuretic peptide (ANP) (A), brain natriuretic peptide (BNP) (B), c-fos (C), and c-jun (D) mRNA expressions, as shown by RT-PCR. Data represent mean ± SEM. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (**P < 0.01).

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  Figure 10. The effects of tannic acid (TA) treatment on nuclear factor-κB (NF-κB) (p65), p38, and toll-like receptor 4 (TLR4) expression, as shown by Western blotting (WB) analysis. Quantification of the NF-κB (p65) (A, B), p38 (C, D) and TLR4 (E, F) protein expression normalization to the β-actin. The mean ± SEM are shown. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (*P < 0.05 and **P < 0.01).

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  Figure 11. The effects of tannic acid (TA) treatment on phosphorylated nuclear factor-κB (NF-κB) (p65), and p38 expression, as shown by Western blotting (WB) analysis. Quantification of the phosphorylated p38 (A, B), and NF-κB (p65) (C, D) protein expression normalization to the β-actin. The mean ± SEM are shown. Compared to the CONT group (##P < 0.01). Compared to the isoproterenol (ISO) group (*P < 0.05 and **P < 0.01).