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Ozone (O 3 ) as a major air pollutant is widely recognized for causing pathological changes of the airway system. However, it is not clear whether O 3 exposure of bronchial epithelial cells (BECs) influences the proliferation and collagen synthesis of submucosal fibroblasts and contributes to the pathogenesis of airway remodeling in diseases, including asthma. In the present study, a co-culture method was applied to culture human lung fibroblasts (HLFs) with human bronchial epithelial cells (HBECs) that were pre-stimulated with O 3 . Following co-culture for up to 24 h, the proliferation of HLFs was measured using MTT colorimetry. Furthermore, the collagen synthesis capacity of HLFs was determined by the level of hydroxyproline. In addition, the protein expression levels of cytokines, including transforming growth factor (TGF)-β1, tumor necrosis factor (TNF)-α and prostaglandin E2 (PGE2) were assessed. Results indicated that the proliferation of HLFs co-cultured with HBECs was significantly inhibited when compared with HLFs cultured alone (P&lt;0.05). By contrast, co-culture with O 3 -stimulated HBECs significantly promoted the proliferation of HLFs compared with the HLFs cultured alone or those cultured with HBECs but no O 3 stimulation, respectively (P&lt;0.05 and P&lt;0.01). Furthermore, similar effects were observed regarding the collagen synthesis capacity of HLFs co-cultured with HBECs for 24. In the supernatant, TGF-β1 concentration was continuously increased over 24 h, whereas the concentration of PGE2 increased and plateaued between 12 to 24 h and TNF-α concentration was not significantly altered during the assessed time period. To conclude, the present results suggest that O 3 pre-exposure of HBECs may promote the transformation of HLFs from the typical inhibitory state into a promoting state with respect to proliferation and collagen synthesis, which may likely occur through a mechanism that influences the balance between pro- and anti-inflammatory factors, including TGF-β1 and PGE2. The present findings may improve the understanding of the mechanism involved in O 3 -induced airway remodeling from a novel perspective of maintenance/loss of steady-state function of the airway epithelium.

Effects of ozone stimulation of bronchial epithelial cells on proliferation and collagen synthesis of co‑cultured lung fibroblasts