Fig 1.
Rosuvastatin treatment on inflammatory infiltration in the lungs.
(A) The illustration shows the establishment of the OVA-mediated chronic allergic asthma model in mice and the groups with or without PAC treatment. (B) The representative image of a lung tissue section stained with Hematoxylin and Eosin (HE) from each experimental group. (C) The blinded inflammation score obtained from the analysis of HE-stained sections. (C and D) The airway wall thickness (C) and smooth muscle thickness (D) from the HE-stained lung sections using Image-Pro Plus software. All images were captured at a magnification of ×200. The data are expressed as the mean ± SD (n = 5) and analyzed using one-way ANOVA followed by Tukey’s post-hoc test. Statistical significance is denoted as *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Fig 2.
Rosuvastatin treatment mitigates airway remodeling in the histology.
(A) Representative images of lung sections stained with PAS (upper) and Masson’s trichrome (down) from each group. (B) Blinded scoring of the percentage of PAS-positive epithelial cells. (C) Blinded quantitative analyses of the area of peribronchial Masson’s trichrome staining using Image-Pro Plus. All images were captured at a magnification of ×200. The data are presented as the mean ± SD (n = 5) and were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. Statistical significance is indicated as *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Fig 3.
Rosuvastatin treatment exhibits anti-fibrotic effects.
(A) Representative images displaying the immunoreactivity of α-SMA, TGF-β, and MMP-9 in lung tissues. (B–D) Quantitative analysis of α-SMA (B), TGF-β (C), and MMP-9 (D) immunohistochemical integral optical density (IOD) using Image-Pro Plus. The images were captured at a magnification of ×200. The data is presented as the mean ± SD (n = 5) and was analyzed using one-way ANOVA, followed by Tukey’s post-hoc test. Statistical significance is denoted as *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Fig 4.
Rosuvastatin treatment reduces the concentrations of various cytokines in the BALF.
(A and B) Comparison of total cell numbers (A) and differential cell counts (B) in the BALF. (C-H) The concentrations of cytokines, including IL-4 (C), IL-5 (D), IL-13 (E), TNF-α (F), TGF-β (G), and MMP-9 (H) in the BALF, were detected by ELISA. The data were presented as the mean ± standard deviation (n = 5) and were analyzed using one-way analysis of variance (ANOVA), followed by Tukey’s post-hoc test. Statistical significance is denoted as *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Fig 5.
Rosuvastatin treatment decreases the levels of OVA-IgE and angiogenesis-related cytokines in the serum.
(A-D) The concentrations of OVA-specific IgE (A), VEGF (B), SDF-1 (C), and CXCR4 (D) in the serum were detected by ELISA. The data were presented as the mean ± standard deviation (n = 5) and were subjected to one-way analysis of variance (ANOVA), followed by Tukey’s post-hoc test. Statistical significance is denoted as *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Fig 6.
Effects of rosuvastatin treatment on AMPK activity and angiogenesis.
(A and B) Representative images (A) and blinded quantitative analyses (B) of p-AMPK levels detected by immunohistochemistry in lung sections. (C) Western blotting and densitometric analyses of p-AMPK and AMPK in lung tissues. (D-G) Immunohistochemical (D) and immunofluorescent (E) staining of lung sections, showing the presence of microvessels, which were further quantified using blinded quantitative analysis of the number (F) and area (G) of microvessels positive for CD31 immunostaining in the sub-epithelial region as well as fluorescence intensities for CD31 staining in the sub-epithelial region (H). Statistical analysis using one-way ANOVA, followed by Tukey’s post-hoc test showed that the results were statistically significant (P < 0.05, P < 0.01, P < 0.001, and P < 0.0001).