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Gallelli, L.; Pelaia, G.; Fratto, D.; Muto, V.; Falcone, D.; Vatrella, A.; Curto, L.S.; Renda, T.; Busceti, M.T.; Liberto, M.C.; Savino, R.; Cazzola, M.; Marsico, S.A.; Maselli, R.
International journal of immunopathology and pharmacology, 04/2010, Letnik: 23, Številka: 2Journal Article
Non-typeable Haemophilus influenzae (NTHi) is one of the most frequently involved pathogens in bacterial exacerbations of chronic obstructive pulmonary disease (COPD). In the airways, the main tissue target of NTHi is bronchial epithelium, where this pathogen can further amplify the inflammatory and structural changes induced by proinflammatory cytokines such as tumour necrosis factor-α (TNF-α). Therefore, the aim of this study is to investigate, in primary cultures of human bronchial epithelial cells, the effects of NTHi on signal transduction pathways, apoptotic events and chemokine production activated by TNF-α. Moreover, we also evaluated the effects exerted on such cellular and molecular phenomena by a corticosteroid drug. p38 mitogen-activated protein kinase (MAPK) phosphorylation was analyzed by Western blotting, using an anti-phospho-p38 MAPK monoclonal antibody. Apoptosis was assayed by active caspase-3 expression. Interleukin-8 (IL-8/CXCL8) was detected in cell-free culture supernatants by ELISA. TNF-α induced a significant increase in p38 MAPK phosphorylation. NTHi was able to potentiate the stimulatory actions of TNF-α on caspase-3 expression and, to a lesser extent, on IL-8 secretion. These effects were significantly (P < 0.01) inhibited by a pharmacological pre-treatment with budesonide. These results suggest that TNF-a is able to stimulate, via activation of p38 MAPK signalling pathway, IL-8 release and airway epithelial cell apoptosis; the latter effect can be markedly potentiated by NTHi. Furthermore, budesonide can be very effective in preventing, through inhibition of p38 MAPK phosphorylation, both structural and proinflammatory changes elicited in bronchial epithelium by TNF-α and NTHi.
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