TY - JOUR
T1 - H2O2 stimulates cystic fibrosis transmembrane conductance regulator through an autocrine prostaglandin pathway, using multidrug-resistant protein-4
AU - Conner, Gregory E.
AU - Ivonnet, Pedro
AU - Gelin, Murline
AU - Whitney, Philip
AU - Salathe, Matthias
PY - 2013/10
Y1 - 2013/10
N2 - Cystic fibrosis transmembrane conductance regulator (CFTR) activity is essential for the maintenance of airway surface liquid depth, and therefore mucociliary clearance. Reactive oxygen species, increased during inflammatory airway diseases, alter CFTR activity. Here, H2O2 levels in the surface liquid ofnormalhumanbronchial epithelial cultures differentiated at the air-liquid interface were estimated, and H2O2- mediated changes in CFTR activitywere examined. In Ussing chambers, H 2O2-induced anion currents were sensitive to the CFTR inhibitors CFTRinh172 and GlyH-101. These currents were absent in cells from patients with cystic fibrosis. Responses to greater than 500 μM H2O2 were transient. Cyclooxygenase inhibitors blocked the H2O2 response, as did EP1 and EP4 receptor antagonists. Amultidrug-resistant protein (MRP) inhibitor and short hairpin RNA directed againstMRP4 blocked H2O2 responses. EP1 and EP4 agonists mimicked H2O2 in both control andMRP4 knockdown cells. Thus, H2O2 activates the synthesis, export, and binding of prostanoids via EP4 and, interestingly, EP1 receptors in normal, differentiated human airway epithelial cells to activate cyclic adenosine monophosphate pathways that in turn activate CFTR channels in the apicalmembrane.
AB - Cystic fibrosis transmembrane conductance regulator (CFTR) activity is essential for the maintenance of airway surface liquid depth, and therefore mucociliary clearance. Reactive oxygen species, increased during inflammatory airway diseases, alter CFTR activity. Here, H2O2 levels in the surface liquid ofnormalhumanbronchial epithelial cultures differentiated at the air-liquid interface were estimated, and H2O2- mediated changes in CFTR activitywere examined. In Ussing chambers, H 2O2-induced anion currents were sensitive to the CFTR inhibitors CFTRinh172 and GlyH-101. These currents were absent in cells from patients with cystic fibrosis. Responses to greater than 500 μM H2O2 were transient. Cyclooxygenase inhibitors blocked the H2O2 response, as did EP1 and EP4 receptor antagonists. Amultidrug-resistant protein (MRP) inhibitor and short hairpin RNA directed againstMRP4 blocked H2O2 responses. EP1 and EP4 agonists mimicked H2O2 in both control andMRP4 knockdown cells. Thus, H2O2 activates the synthesis, export, and binding of prostanoids via EP4 and, interestingly, EP1 receptors in normal, differentiated human airway epithelial cells to activate cyclic adenosine monophosphate pathways that in turn activate CFTR channels in the apicalmembrane.
KW - CFTR
KW - EP1
KW - EP4
KW - HO
KW - MRP4
UR - http://www.scopus.com/inward/record.url?scp=84886564112&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84886564112&partnerID=8YFLogxK
U2 - 10.1165/rcmb.2013-0156OC
DO - 10.1165/rcmb.2013-0156OC
M3 - Article
C2 - 23742099
AN - SCOPUS:84886564112
VL - 49
SP - 672
EP - 679
JO - American Journal of Respiratory Cell and Molecular Biology
JF - American Journal of Respiratory Cell and Molecular Biology
SN - 1044-1549
IS - 4
ER -