Autocrine Acetylcholine, Induced by IL-17A via NFκB and ERK1/2 Pathway Activation, Promotes MUC5AC and IL-8 Synthesis in Bronchial Epithelial Cells

Figure 5

Effects of PD098,059 (25 μM), Bay11-7082 (50 μM), and Tiotropium (100 nM) on ACh in 16-HBE cells. Cells were preincubated for 1 h with PD098,059 (25 μM) or Bay11-7082 (50 μM) and then stimulated with rhIL-17A 20 ng/mL for 24 h to evaluate (a) ACh binding by flow cytometry. Bars represent mean ± SD of fluorescence mean intensity (FMI). (b) Representative flow cytometry is shown. (c) ACh production, expressed as pmoli/μg protein. Bars represent mean ± SD of three different experiments. Then, 16-HBE cells were preincubated for 1 h with Tiotropium (100 nM) and then stimulated with rhIL-17A 20 ng/mL for 24 h to evaluate (d) ACh binding by flow cytometry. Bars represent mean ± SD of fluorescence mean intensity (FMI). (e) Representative flow cytometry is shown. Statistical analysis was performed by ANOVA test followed by Fisher’s PLSD multiple comparison test. The black curve represents the anti-IgG isotype negative control antibody.

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