Simvastatin Reduces NETosis to Attenuate Severe Asthma by Inhibiting PAD4 Expression

Figure 3

Adoptively transferring neutrophils from OVA+LPS mice to simvastatin-treated OVA+LPS mice was sufficient to trigger the TH17-mediated neutrophilic asthma. (a) Confocal microscopy staining of CitH3⁺ MPO⁺ DAPI⁺ NETs released from ex vivo-cultured neutrophils and the lung sections of the indicated groups of mice. (b) Representative blots of CitH3 and actin (loading control) assessed by Western blot of lung protein extracts from mice. (c) Quantification of normalized CitH3 levels in lung protein extracts of mice. (d, f) Representative hematoxylin and eosin (H&E) staining of lung sections of mice. (e) Representative PAS staining of lung sections of mice. (g) Quantification of PAS-stained epithelial cells per bronchi showing airway mucus production in mice. (h, i) Total cell counts, eosinophil counts, and neutrophil counts (10⁵ cells/ml) in the BALF. (j, k) Levels of IL-1β, IFN-γ, IL-4, and IL-17 in the BALF. (l–o) Flow cytometric analysis of percentages of TH2, TH17, and Treg lymphocytes in the lung.