Eicosanoid Release Is Increased by Membrane Destabilization and CFTR Inhibition in Calu-3 Cells 英文参考文献.docVIP

EicosanoidReleaseIsIncreasedbyMembrane DestabilizationandCFTRInhibitioninCalu-3Cells FlorenceBorot1,Diane-LoreVieu1.,GrazynaFaure2.,JanineFritsch1,JulienColas1,SandraMoriceau1, MaryvonneBaudouin-Legros1,FranckBrouillard1,JesusAyala-Sanmartin3,LhousseineTouqui4 ,Marc Chanson5,AleksanderEdelman1,MarioOllero1* 1INSERM,U845,Universite′ParisDescartes,Faculte′ deMe′decineParisDescartes,Paris,France,2InstitutPasteur,Unite′d’ImmunologieStructurale,CNRS,URA2185,Paris, France,3CNRS,UMR7203,GroupeN.J.Conte′,LaboratoiredesBioMole′cules,Paris,France,4InstitutPasteur,Unite′deDe′fenseInne′eetInflammation,INSERM,U874,Paris, France,5Laboratoired’InvestigationCliniqueIII,Ho?pitauxUniversitairesetFaculte′ deMe′decine, Gene`ve,Switzerland Abstract The antiinflammatory protein annexin-1 (ANXA1) and the adaptor S100A10 (p11), inhibit cytosolic phospholipase A2 (cPLA2a) by direct interaction. Since the latter is responsible for the cleavage of arachidonic acid at membrane phospholipids, all three proteins modulate eicosanoid production. We have previously shown the association of ANXA1 expression with that of CFTR, the multifactorial protein mutated in cystic fibrosis. This could in part account for the abnormal inflammatory status characteristic of this disease. We postulated that CFTR participates in the regulation of eicosanoidreleasebydirectinteractionwithacomplexcontainingANXA1,p11andcPLA2a.Wefirstanalyzedbyplasmon surfaceresonancetheinvitrobindingofCFTRtothethreeproteins.Asignificantinteractionbetweenp11andtheNBD1 domainofCFTRwasfound.WeobservedinCalu-3cellsarapidandpartialredistributionofallfourproteinsindetergent resistantmembranes(DRM)inducedbyTNF-a.ThiswasconcomitantwithincreasedIL-8synthesisandcPLA2aactivation, ultimatelyresultingineicosanoid(PGE2andLTB4)overproduction.DRMdestabilizingagentmethyl-b-cyclodextrininduced further cPLA2a activation and eicosanoid release, but inhibited IL-8 synthesis. We tested in parallel the effect of short exposureofcellstoCFTRinhibitors