Berberine impairs coxsackievirus B3‐induced myocarditis through the inhibition of virus replication and host pro‐inflammatory response

Figures

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  Figure 1. The BBR protects mice from CVB3‐induced viral myocarditis. A, The chemical structure of BBR. B, Experimental protocol. One day before CVB3 infection, the mice were intragastrically administered with BBR for 7 days. On the 7th day, all experiments were measured. C, The survival rates of mouse after CVB3 infection or/and BBR treatment (n = 24). D, The measurement of mouse heart weight to body weight ratio after CVB3 infection or/and BBR treatment (n = 6). E, The representative photographs of the whole hearts. Data show mean ± SEM of mice. BBR, berberine; CVB3, coxsackievirus B3. ***p＜.001, n.s., no significant

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  Figure 2. BBR protects mice heart lesion from CVB3‐induced myocarditis mice. A, Heart histological examination by H&E staining. Upper images, scale bars = 50 μm, magnification ×200 and lower images, scale bars = 20 μm, magnification ×400. B, The heart lesion degree evaluated by blind scoring methods from H&E staining (n = 8). C, The myocardial zymogram containing AST, (D) CK‐MB, (E) LDH, and (F) CK in mice serum were determined by an automatic biochemistry analyzer (n = 6). Data show mean values ± SEM of mice. AST, aspartate aminotransferase; BBR, berberine; CK, creatine kinase; CK‐MB, creatine kinase‐MB fraction; CVB3, coxsackievirus B3; H&E, hematoxylin and eosin; LDH, lactate dehydrogenase. **p＜.01, ***p＜.001

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  Figure 3. BBR protects heart function from CVB3‐induced myocarditis mice. A, Echocardiogram experiments. B‐C, The results of ejection fraction and fractional shortening are shown. Data show mean ± SEM of mice. BBR, berberine; CVB3, coxsackievirus B3. *p < .05, **p < .01, ***p < .001

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  Figure 4. BBR inhibits titer of CVB3 in vivo. A, The IHC staining for the CVB3 particles in mouse myocardium of the indicated groups. B, The mRNA expression level of the virus vp1 gene in mouse myocardium upon CVB3 infection or BBR treatment (n = 3). C, The viral titer from mouse hearts was determined by the Reed‐Muench method (n = 3). Data show mean ± SEM of mice. BBR, berberine; CVB3, coxsackievirus B3; IHC, immunohistochemistry; mrna, messenger RNA. *p < .05, ***p < .001

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  Figure 5. BBR diminishes CVB3‐induced expression of cytokines and chemokines. The production levels of (A) CCL2, (B) CCL5, (C) CXCL10, (D) TNF‐α, (E) IL‐6, and (F) IL‐1β in mice serum were determined by ELISA (n = 4). Data show mean ± SEM of mice. BBR, berberine; CCL2, C‐C motif chemokine ligand 2; CVB3, coxsackievirus B3; CXCL10, C‐X‐C motif chemokine ligand 10; IL‐6, interleukin‐6; TNF‐α, tumor necrosis factor‐α. *p < .05, **p＜.01, ***p＜.001

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  Figure 6. BBR attenuates the increased infiltration of immune cells in CVB3‐infected mice. A, The infiltration of CD68+ macrophages were evaluated by immunofluorescence staining. B–E, The proportion of CD4+ and CD8+ T lymphocytes to total cells isolated from mouse spleens was determined by flow cytometry (n = 3). BBR, berberine; CVB3, coxsackievirus B3