Inhibition of pyocyanin-potentiated IL-8 release by steroids in bronchial epithelial cells

• Published in: Respiratory medicine 2006-09, Vol.100 (9), p.1614-1622
• Main Authors: Pan, Nin Y., Hui, Wing S., Tipoe, George L., Taylor, Graham W., Leung, Raymond Y.H., Lam, Wah K., Tsang, Kenneth W.T., Mak, Judith C.W.
• Format: Article
• Language: English
• Subjects: Androstadienes - pharmacology; Anti-Inflammatory Agents - pharmacology; Biological and medical sciences; Bones, joints and connective tissue. Antiinflammatory agents; Bronchi - cytology; Bronchi - drug effects; Bronchi - metabolism; Bronchiectasis; Budesonide - pharmacology; Cytokines; Dexamethasone; Dexamethasone - pharmacology; Disease; Enzyme-Linked Immunosorbent Assay; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Fluticasone; Glucocorticoids - pharmacology; Humans; Interleukin-1 - pharmacology; Interleukin-8; Interleukin-8 - secretion; Kinases; Medical sciences; Pathogenesis; Pharmacology. Drug treatments; Phorbol 12,13-Dibutyrate - pharmacology; Pneumology; Polymerase Chain Reaction; Protein Kinase C - drug effects; Protein Kinase C - metabolism; Proteins; Pseudomonas aeruginosa pyocyanin; Pyocyanine - pharmacology; Respiratory system : syndromes and miscellaneous diseases; RNA, Messenger - drug effects; RNA, Messenger - metabolism; Steroids
• ISSN: 0954-6111; 1532-3064
• DOI: 10.1016/j.rmed.2005.12.003

Airway epithelial cells are the first targets of environmental stimuli and local cytokines. Pyocyanin-induced synergism with interleukin (IL)-1 or tumour necrosis factor (TNF) in triggering IL-8 release has been documented previously. In this study, IL-8 mRNA and protein expression were examined in cultured human bronchial epithelial cells (BEAS-2B) stimulated with pyocyanin alone, and in combination with IL-1 β or phorbol 12,13-dibutyrate (PDBu) in the absence and presence of a group of glucocorticoids. IL-8 mRNA was measured by RT-PCR, and IL-8 protein by ELISA (cell supernatants). Pyocyanin alone produced no increase in IL-8 mRNA and release. However, pyocyanin upregulated the stimulatory effect of IL-1 β or PDBu on the release of IL-8 in a dose-dependent manner. The stimulatory effect of pyocyanin on the IL-1 β- or PDBu-stimulated IL-8 release was reduced in the presence of dexamethasone, budesonide, and fluticasone. Budesonide and fluticasone were 10-fold more potent than dexamethasone. The protein kinase C (PKC) inhibitor, Go6976, also significantly reduced the stimulatory effect of pyocyanin on IL-1 β, and PDBu increased IL-8 release. In conclusion, this study shows that PKC signal pathway seems to be involved in the pyocyanin-mediated upregulation of the IL-1 β and PDBu-induced IL-8 release in BEAS-2B cells. These findings suggest that a vicious cycle perpetuating inflammation may exist in the biologic milieu of bronchiectatic patients infected with Pseudomonas aeruginosa due to the production of pyocyanin. The priming action of pyocyanin appears to be blocked by glucocorticoids, thus providing in vitro data in support of the clinical efficacy of inhaled glucocorticoids as anti-inflammatory drugs.