Berberine plays a cardioprotective role by inhibiting macrophage Wnt5a/β‐catenin pathway in the myocardium of mice after myocardial infarction

Figures

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  FIGURE 1. Berberine can improve cardiac function and inhibit ventricular remodeling after Myocardial infarction (MI) in mice. (a) Representative echocardiography images of sham group, MI group, and Berberine group. (b) left ventricular diastolic dimension, left ventricular internal dimension systole, left ventricular ejection fraction, and left ventricular fraction shortening were measured by echocardiography. (c) Body weight changes of each group. (d) The ratio of heart weight to body weight of mice in each group. (e) Representative Masson staining (Bar = 1 mm above, Bar = 200um below) sections and (f) the percentage of infarct region fibrosis of mice in each group. n = 6 animals per group. Data are presented as mean ± SEM. *p < .05, **p < .01, ***p < .001

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  FIGURE 2. Berberine plays a cardioprotective role by mediating the immune cells and cytokines in mice with Myocardial infarction (MI). (a) Flow cytometry gating strategies for each cell population. (b) The bar charts show the percentages of neutrophils, Macrophages, Ly6Chigh macrophages, Ly6Clow macrophages, DCs, (c) and CD4+ T cells, Foxp3+CD4+ Treg cells, ROR‐γt+CD4+ T, and CD8+ T cells by flow cytometry. (d) The mRNA expression of TNF‐α, IL‐6, IL‐18, MIP‐1β, TGF‐β, IL‐10, VEGF‐α by RT‐PCR, using infarcted heart tissues from the sham group, MI group, and Berberine group. n = 6 animals per group. Data are presented as mean ± SEM. *p < .05, **p < .01, ***p < .001

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  FIGURE 3. Berberine can alleviate oxidative damage of H9C2 and improve the migration and angiogenesis ability of HUVEC. H9C2 cells were administrated with H2O2, and low or high dose of Berberine (BBR) under the condition of hypoxia injury, (a) the morphology of the cells was observed, (b) the concentrations of malondialdehyde and superoxide dismutase, (c) the staining of 7‐AAD/Annexin V (left), and the percentages of early apoptotic cells (right) were measured in these cells. (d) The migration ability of HUVECs was assayed (left) and the migration area was calculated (right) under the condition of hypoxia injury and the treatment of H2O2 and different concentration of BBR. Data are presented as mean ± SEM. *p < .05, **p < .01, ***p < .001

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  FIGURE 4. Berberine alleviates MI in mice by inhibiting Wnt5a pathway. (a‐b) Infarcted heart tissues from sham, Myocardial infarction (MI), and Berberine (BBR) groups were collected for protein and mRNA preparation. The protein levels of Wnt5a and β‐catenin and mRNA level of Wnt5a were measured by western blotting and PCR, respectively. Lymphocytes were prepared from heart tissues of three groups, and cultured for macrophage selection. (c) The extracted cardiac macrophages were evaluated by flow cytometry, (d) and the protein levels of Wnt5a and β‐catenin were assayed by western blotting. The effect of cardiac macrophages on angiogenesis was evaluated by tube formation experiment co‐cultured with supernatant of cultured macrophages. (e) The images and numbers of formed tubes were examined. (f) The pictures show the representative echocardiography images, and the bar charts indicate the levels of left ventricular diastolic dimension, left ventricular internal dimension systole, left ventricular ejection fraction, and left ventricular fraction shortening from sham, MI, BBR, and FH535 groups. n = 5 animals per group. Data are presented as mean ± SEM. *p < .05, **p < .01, ***p < .001

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  FIGURE 5. Schematic depiction of alteration of the immune homeostasis after Berberine (BBR) application in Myocardial infarction. Mechanism diagram of BBR regulating the immunity of mice after myocardial infarction by inhibiting Wnt5a