06). The co-exposure to cigarette smoke did not increase IL-5 levels in the lung tissue or the number of IL-5 positive cells in the peribronchovascular space (Fig. 4C and D, respectively). The OVA groups showed a significant increase C59 in IL-5 levels in the lung tissue when compared with all of the other groups (p = 0.004); however, this difference could not be detected in the peribronchovascular
space, despite graphic similarities (p = 0.06). Cigarette smoke exposure did not increase eotaxin levels in the lung tissue (Fig. 4E). The OVA group showed a significant increase in eotaxin when compared with all of the other groups (p = 0.01). In contrast, an increase in IFN-γ levels in the lung tissue was observed in the OVA + CS group when compared with all of the other groups (p = 0.001) ( Fig. 4F). Fig. 5 shows a panel with the levels of IL-10 measured in the Bio-Plex assay and the numbers of IL-10-positive
cells in the Kinase Inhibitor Library purchase bronchial epithelium (Fig. 5A and B, respectively). There was an increase in IL-10 levels in the CS, OVA and OVA + CS groups, with the OVA + CS group significantly different from all of the other groups (p = 0.001). The CS and OVA groups also showed significant differences compared with the Control group (p < 0.05) ( Fig. 5A). The abundance of IL-10-positive cells was also increased in the groups exposed to cigarette smoke when compared with the Control group (p < 0.05) ( Fig. 5B). Exposure to ovalbumin
resulted in a non-significant increase in collagen fiber content in the peribronchovascular area (p = 0.06 compared with the control group, Fig. 6). Only the OVA + CS group showed a significant increase of collagen fiber content in the peribronchovascular area (p = 0.001 compared with the other three groups). Panels A–D show representative photomicrographs of collagen content in the bronchovascular structures in the four experimental groups following staining G protein-coupled receptor kinase for collagen fibers. The OVA + CS group showed a significant increase in the abundance of TGF-β-positive cells in the bronchial epithelium (p < 0.005 compared with the Control and CS groups, Fig. 7A). Isolated exposure to either OVA or cigarette smoke did not increase the density of TGF-β-positive cells in the epithelium. In addition, there was a strong correlation between TGF-β-positive epithelial cells and peribronchovascular collagen fiber content ( Fig. 6) in the OVA + CS group (r = 0.91; p = 0.01). The cytokine assay also showed a significant increase in GM-CSF levels in the OVA + CS group compared with all of the other groups (p = 0.004) ( Fig. 7B). Cigarette smoke exposure also increased VEGF levels, as indicated in Fig. 7C. The OVA + CS group showed a significant difference in VEGF levels compared with the Control and OVA groups (p = 0.03). The CS group showed a similar increase in VEGF levels when compared with the control mice (p = 0.01).