Ex Vivo Effect of Varespladib on Secretory Phospholipase A2 Alveolar Activity in Infants with ARDS 英文参考文献.docVIP

ExVivoEffectofVarespladibonSecretoryPhospholipase A2AlveolarActivityinInfantswithARDS DanieleDeLuca1,2*,AngeloMinucci1,MarcoPiastra2,PaolaE.Cogo3,FrancescaVendittelli1, LauraMarzano2,LeonardaGentile1,BrunoGiardina1,GiorgioConti2,EttoreD.Capoluongo1 1LaboratoryofClinicalMolecularBiology,DepartmentofBiochemistry,UniversityHospital‘‘A.Gemelli’’,CatholicUniversityoftheSacredHeart,Rome,Italy,2Pediatric IntensiveCareUnit,DepartmentofAnaesthesiologyandIntensiveCare,UniversityHospital‘‘A.Gemelli’’,CatholicUniversityoftheSacredHeart,Rome,Italy,3Pediatric IntensiveCareUnit,DepartmentofPediatrics,HospitalandUniversityofPadua,Padua,Italy Abstract Background:SecretoryphospholipaseA2(sPLA2)playsapivotalroleinacuterespiratorydistresssyndrome(ARDS).This enzyme seems an interesting target to reduce surfactant catabolism and lung tissue inflammation. Varespladib is a specificallydesignedindolicsPLA2inhibitor,whichhasshownpromisingresultsinanimalsandadults.Nospecificdatain pediatricARDSpatientsareyetavailable. Methods: We studied varespladib in broncho-alveolar lavage (BAL) fluids obtained ex vivofrom pediatric ARDS patients. ClinicaldataandworstgasexchangevaluesduringtheARDScoursewererecorded.Samplesweretreatedwithsalineor 10–40–100mM varespladib and incubated at 37uC. Total sPLA2 activity was measured by non-radioactive method. BAL samples were subjected to western blotting to identify the main sPLA isotypes with different sensitivity to varespladib. Resultswascorrectedforlavagedilutionusingtheserum-to-BALurearatioandforvarespladibabsorbance. Results:VarespladibreducessPLA2activity(p,0.0001)at10,40and100mM;bothsPLA2activityreductionanditsratioto total proteins significantly raise with increasing varespladib concentrations (p,0.001). IC50 was 80mM. Western blotting revealedthepresenceofsPLA2-IIAand–IBisotypesinBALsamples.SignificantcorrelationsexistbetweenthesPLA2activity reduction/proteinsratioandPaO2(rho=0.63;p,0.001),PaO2/FiO2(rho=0.7;p,0.001),oxygenation(rho=20.6;p,0.001) andve