RESUMO
Objective To observe the expressions of peroxisome proliferator activatedγ (PPARγ),IL-17 and calcium activated family member 1 (hCLCA1) mRNA and protein in A549 cell infected with respiratory syncytial virus(RSV),and to explore the effect of rosiglitazone and anti-IL-17 Ab.Methods A549 cells were seeded in 6-well culture plates and randomly divided into six groups.DMSO/RSV group received medium containing 0.02％ DMSO 30min prior to RSV infection.Rosiglitazone/RSV group received rosiglitazone (101μmol/L) 30 min prior to RSV infection.GW9662/rosiglitazone/RSV group was first exposed to GW9662 (10μmol/L)for 30 min,prior to rosiglitazone and RSV infection.Anti-IL-17 Ab (0.15μmol/L) was administered to anti-IL-17 Ab/RSV group 2 hour prior to RSV infection.Supernatants were harvested at 6h,12h and 24h post-RSV infection,retrospectively.The expression of PPARγ,IL-17,hCLCA1 mRNA were measured by real-time quantitative PCR.Protein levels of hCLCA1 and IL-17 were measured by Western blot and ELISA,retrospectively.Results The levels of PPARγ,IL-17and hCLCA1 mRNA and protein were higher in DMSO/RSV group than those in cell control group (P ＜ 0.05) at three time points.The expression of IL-17,hCLCA1 mRNA and protein were lower both in rosiglitazone/RSV group and anti-IL-17 Ab/RSV group than those in DMSO/RSV group at the same time (P ＜0.05).While,both IL-17,hCLCA1 protein and mRNA expression in GW9662/Rosiglitazone/RSV group were similar to that in DMSO/RSV group (P＜ 0.05).Compared with that at 12h and 24h,rosiglitazone and anti-IL-17 Ab had the strongest inhibition effect onthe expression of IL-17 mRNA and protein at 6h (P ＜ 0.05).Conclusion Rosiglitazone can inhibit RSV-induced expression of IL-17,hCLCA1 and may inhibit airway mucus hypersecretion by increasing the activity of PPARγ.
RESUMO
Objective To observe the expressions of peroxisome proliferator activatedγ (PPARγ),IL-17 and calcium activated family member 1 (hCLCA1) mRNA and protein in A549 cell infected with respiratory syncytial virus(RSV),and to explore the effect of rosiglitazone and anti-IL-17 Ab.Methods A549 cells were seeded in 6-well culture plates and randomly divided into six groups.DMSO/RSV group received medium containing 0.02％ DMSO 30min prior to RSV infection.Rosiglitazone/RSV group received rosiglitazone (101μmol/L) 30 min prior to RSV infection.GW9662/rosiglitazone/RSV group was first exposed to GW9662 (10μmol/L)for 30 min,prior to rosiglitazone and RSV infection.Anti-IL-17 Ab (0.15μmol/L) was administered to anti-IL-17 Ab/RSV group 2 hour prior to RSV infection.Supernatants were harvested at 6h,12h and 24h post-RSV infection,retrospectively.The expression of PPARγ,IL-17,hCLCA1 mRNA were measured by real-time quantitative PCR.Protein levels of hCLCA1 and IL-17 were measured by Western blot and ELISA,retrospectively.Results The levels of PPARγ,IL-17and hCLCA1 mRNA and protein were higher in DMSO/RSV group than those in cell control group (P ＜ 0.05) at three time points.The expression of IL-17,hCLCA1 mRNA and protein were lower both in rosiglitazone/RSV group and anti-IL-17 Ab/RSV group than those in DMSO/RSV group at the same time (P ＜0.05).While,both IL-17,hCLCA1 protein and mRNA expression in GW9662/Rosiglitazone/RSV group were similar to that in DMSO/RSV group (P＜ 0.05).Compared with that at 12h and 24h,rosiglitazone and anti-IL-17 Ab had the strongest inhibition effect onthe expression of IL-17 mRNA and protein at 6h (P ＜ 0.05).Conclusion Rosiglitazone can inhibit RSV-induced expression of IL-17,hCLCA1 and may inhibit airway mucus hypersecretion by increasing the activity of PPARγ.
RESUMO
OBJECTIVES/HYPOTHESIS: Mucus hypersecretion is a hallmarks of chronic rhinosinusitis. The expression of MUC5AC, a major respiratory mucin gene, is increased in chronic rhinosinusitis. The mechanisms inducing mucus hypersecretion have not been fully evaluated in chronic rhinosinusitis. Human Ca2+ -activated Cl- channel 1 (hCLCA1) is implicated in the regulation of mucus production, airway fluid, and electrolyte transport. The present study objectives was to investigate the expression of hCLCA1 in chronic rhinosinusitis and evaluate whether its level is altered by stimulation with type 1 T helper (Th1) and Th2 cytokines, and to determine the possible role of hCLCA1 on the regulation of mucin 5AC (MUC5AC) production. STUDY DESIGN: Controlled prospective study. METHODS: The expression of hCLCA1 and MUC5AC in normal and inflammatory ethmoid mucosa was determined by real-time polymerase chain reaction, immunohistochemistry, and Western blot. In cultured cells, the expression of hCLCA1 and MUC5AC was measured after stimulation with Th1 and Th2 cytokines. In a supernatant, the MUC5AC level was analyzed using enzyme-linked immunosorbent assay after treatment with niflumic acid. RESULTS: The levels of hCLCA1 and MUC5AC were increased in chronic rhinosinusitis, irrespective of nasal polyp presence, where they were distributed in superficial epithelial cells and submucosal glands. In cultured cells treated with interleukin (IL)-9, IL-4, IL-13, tumor necrosis factor-α, transforming growth factor-ß, interferon-γ, and IL-1ß, the expression of hCLCA1 and MUC5AC was increased. In cells treated with niflumic acid, the production of MUC5AC was inhibited. CONCLUSIONS: The current findings indicate that the expression of hCLCA1 is increased in chronic rhinosinusitis and may be regulated by Th1 and Th2 cytokines, possibly contributing to the production of MUC5AC. LEVEL OF EVIDENCE: NA Laryngoscope, 126:E347-E355, 2016.