Cell Viability and Cytokine Production of Human Alveolar Epithelial Cells Following Exposure to Sulphur Dioxide

Exposure to air pollutants is significantly associated with health risks ranging from bronchial reactivity to morbidity and mortality. However, the precise mechanisms are not always fully understood. The aim of this study was to investigate the effects of sulphur dioxide (SO2) on cell viability and cytokine production of A549-human pulmonary epithelial cells. Test atmospheres of SO2 were generated using a direct dilution method and calibrated by ion-chromatography. Test atmospheres were delivered to lung cells cultured on porous membranes (0.4 μm) using Harvard Navicyte horizontal diffusion chamber systems. The cytotoxic endpoints were investigated using the MTS (tetrazolium salt; Promega), NRU (neutral red uptake; Sigma) and ATP (adenosine triphosphate; Promega) assays. Expression of inflammatory markers including tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) were evaluated using double-antibody immunometric assays. Dose-dependent effects of SO2 were observed in A549 cells using all in vitro assays at test concentrations (10-200 ppm). The ATP assay appeared to be the most sensitive test (IC50 = 48 ± 2.83 ppm) that may related to the impaired metabolic activity of the cells following SO2 exposure. After analysis of TNF-a, no statistically significant differences were observed between control and exposed cells. However, the IL-6 production in A549 cells was significantly reduced in a dose-dependent manner (P<0.05). These results suggest that SO2 may induce a functional alteration of cells of the pulmonary epithelial preventing cells to produce adequate amounts of IL-6. IL-6 as a multifunctional proinflammatory cytokine may regulate cellular responses and plays a significant role in inflammation and tissue injury.