Sacubitril/Valsartan Alleviates Experimental Autoimmune Myocarditis by Inhibiting Th17 Cell Differentiation Independently of the NLRP3 Inflammasome Pathway

Figures

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  FIGURE 1. Sac/Val attenuated the severity of EAM. (A) HW/BW in each treatment group. (B) Typical histopathologic pictures of myocardial tissues (H and E staining, × 200). (C) The statistical graph of pathological scores in each treatment group. (D) Levels of serum cTnT in each treatment group. (E) Levels of serum hsCRP in each treatment group. (F) Levels of serum IL-6 in each treatment group. All experiments had six mice per group and data were expressed as means ± SEM and analysed by one-way ANOVA. * p < 0.05 vs. Control group; # p < 0.05 vs. EAM group; $ p < 0.05 vs. Valsartan group.

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  FIGURE 2. Sac/Val had no effect on activation of NLRP3 inflammasomes. (A) Representative western blotting results for cardiac NLRP3, ASC, and Caspase-1. (B) Statistical analysis of the western blotting results. (C) Representative IHC images of cardiac IL-1β and IL-18 (magnification × 400). (D) Statistical analysis of the IHC results. All experiments had six mice per group and data were expressed as means ± SEM and analysed by one-way ANOVA. * p < 0.05 vs. Control group.

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  FIGURE 3. Sac/Val inhibited the differentiation of Th17 cells in myocardial tissue. (A) Representative IHC images of cardiac IFN-γ and IL-17 (magnification × 400). (B) Semiquantitative analysis of the IHC results. (C) Levels of peripheral blood IFN-γ and IL-17 in each treatment group. (D) Representative flow cytometry images of Th1 (IFN-γ+) and Th17 (IL-17+) cells gated on CD4+ T cells. (E) Statistical analysis of the flow cytometry results. All experiments had six mice per group and data were expressed as means ± SEM and analysed by one-way ANOVA. * p < 0.05 versus Control group; # p < 0.05 vs. EAM group; $ p < 0.05 vs. Valsartan group.

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  FIGURE 4. Sac/Val lowered levels of RORγt in myocardial tissue. (A) Representative western blotting results for cardiac T-bet and RORγt. (B) The statistical graph of the western blotting results. All experiments had six mice per group and data were expressed as means ± SEM and analysed by one-way ANOVA. * p < 0.05 vs. Control group; # p < 0.05 vs. EAM group; $ p < 0.05 vs. Valsartan group.

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  FIGURE 5. sGC and NF-κB p65 functioned as signaling molecules. (A) Representative western blotting images and statistical analysis for cardiac sGC-α1 and sGC-β1. (B) Representative western blotting images and statistical analysis for cardiac phospho-/total-NF-κB p65 signaling pathway. All experiments had six mice per group and data were expressed as means ± SEM and analysed by one-way ANOVA. * p < 0.05 vs. Control group; # p < 0.05 vs. EAM group; $ p < 0.05 vs. Valsartan group.