Coenzyme Q10 mitigates macrophage mediated inflammation in heart following myocardial infarction via the NLRP3/IL1β pathway

Figures

• 

  Fig. 1. Plasma levels of CoQ10 are lower in MI patients (administrated with stains) compared to those in healthy controls, and supplementation with CoQ10 accelerates the recovery of cardiac function in MI patients after PCI therapy. (A) Illustration of the study design of the clinical experiment. (B) The baseline concentration of plasma CoQ10 of healthy controls and MI patients were analyzed by liquid chromatographic-tandem mass spectra-metric (LC-MS/MS) (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). (C) Sequential echocardiographic analyses showed that MI patients supplemented with CoQ10 display a better recovery of cardiac function at 1-month and 3-month post-PCI compared with MI patients without CoQ10 treatment (Statistical analysis was performed using a Mann-Whitney U test). (D) The LnBNP of patients in the MI group and MI + CoQ10 group at 3-month post-PCI demonstrated a greater reduction in MI patients supplemented with CoQ10 compared with MI patients without CoQ10 treatment (Statistical analysis was performed using a Mann-Whitney U test). Grey indicates the control group (healthy volunteers); Red indicates the blank control group (MI patients);…

• 

  Fig. 2. Mice with CoQ10 treatment have a better survival rate and attenuated cardiac remodeling after LAD ligation. (A) Kaplan-Meier survival curve of mice administrated with Vehicle or CoQ10 following LAD ligation (Statistical analysis was performed using the log-rank test). (B)(C) Representative echocardiographic images and analyses of mice at 28 days after sham or LAD ligation surgeries. Mice were administered with either vehicle or CoQ10 (Statistical analyses were performed using 2-way ANOVA with Tukey’s multiple comparison tests). (D)(E) Representative images and analysis of WGA staining of the heart sections from mice of each group at 28 days after sham or LAD ligation surgeries, administrated with vehicle or CoQ10. 200×, scale bars indicate 100 μm (Statistical analysis was performed using 2-way ANOVA with Tukey’s multiple comparison tests). (F)(G) Representative images and analysis of Masson’s trichrome staining of the heart tissue sections from mice of each group at 28 days after sham or LAD ligation surgeries, administrated with vehicle or CoQ10. Scale bars indicate 1 mm (Statistical analysis was performed using 2-way ANOVA with Sidak’s multiple comparison tests). (H) Serum BNP…

• 

  Fig. 3. CoQ10 inhibits the ROS generation and activation of NLRP3/IL1β signaling in macrophages. (A) Gene expression analysis in peritoneal macrophages with or without 6-hour stimulation of LPS/INFγ, in the presence or absence of CoQ10 (Statistical analysis was performed using 2-way ANOVA with Sidak’s multiple comparison tests). (B) Heatmap of RNA script reads of specific genes related to inflammatory cytokines, inflammatory chemokines, macrophage activation, NOD-like receptor signaling, coenzyme metabolic process, fatty acid beta oxidation, ROS biosynthetic process between LPS/IFNγ-primed macrophages treated with or without CoQ10. (C) KEGG enrichment analysis of top-ranked pathways of differentially expressed genes between LPS/IFNγ-primed macrophages treated with or without CoQ10. (D) Western blot analysis of intracellular NLRP3 in peritoneal macrophages after 6 h of stimulation with LPS/INFγ in the presence of vehicle or CoQ10 (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). Data were normalized to β-actin. (E) Western blot analysis of intracellular pro-IL1β in the macrophages after 6 h of stimulation with LPS/INFγ in the presence of vehicle or CoQ10…

• 

  Fig. 4. IL1β production-related post-infarction cardiac inflammation was suppressed by CoQ10 treatment. (A) Analysis of gene expression in the infarct myocardium from each group of mice at 3 days after sham/LAD-ligation surgeries (Statistical analysis was performed using 2-way ANOVA with Sidak’s multiple comparison tests). Mice were treated with either vehicle or CoQ10. (B)(C) Representative images and analysis of the IL1β immunofluorescence staining in the Mac3-positive macrophages from mice treated with vehicle or CoQ10 at 28 days after LAD ligation (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). 630×. Scale bars indicate 20 μm. (D) Representative flow cytometry analysis of left ventricles after sham/LAD ligation surgeries. Cells were defined as neutrophils (Neut, CD45.2+CD64−Ly6G+), monocytes (Mono, CD45.2+Ly6G−CD64intLy6Chi), macrophages (Mac, CD45.2+Ly6G−CD64+Ly6Clo), or CCR2+ macrophages (CCR2+Mac, CD45.2+Ly6G−CD64+Ly6CloCCR2+). (E) Quantification of the mean fluorescence intensity (MFI) of IL-1β in the CCR2+ macrophages of mice 3 days after LAD ligation, either treated with vehicle or CoQ10 (Statistical analysis was performed using a two-tailed…