Effects of budesonide on P38 MAPK activation, apoptosis and IL-8 secretion, induced by TNF-alpha and Haemophilus influenzae in human bronchial epithelial cells.
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-alpha (TNF-alpha). 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-alpha. 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-alpha induced a significant increase in p38 MAPK phosphorylation. NTHi was able to potentiate the stimulatory actions of TNF-alpha on caspase-3 expression and, to a lesser extent, on IL-8 secretion. These effects were significantly (P less than 0.01) inhibited by a pharmacological pre-treatment with budesonide. These results suggest that TNF-alpha 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-alpha and NTHi.
Gallelli
L
L
Department of Experimental and Clinical Medicine, University Magna Grecia of Catanzaro, Catanzaro, Italy.
Pelaia
G
G
Fratto
D
D
Muto
V
V
Falcone
D
D
Vatrella
A
A
Curto
L S
LS
Renda
T
T
Busceti
M T
MT
Liberto
M C
MC
Savino
R
R
Cazzola
M
M
Marsico
S A
SA
Maselli
R
R
D017209:Apoptosis / Q000187:drug effects*; D001980:Bronchi / Q000378:metabolism*; D019819:Budesonide / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism; D006193:Haemophilus influenzae / Q000502:physiology*; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis*; D010766:Phosphorylation; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D019819:Budesonide; D048051:p38 Mitogen-Activated Protein Kinases; D053148:Caspase 3