Flagellin-Induced Corneal Antimicrobial Peptide Production and Wound Repair Involve a Novel NF-κB–Independent and EGFR-Dependent Pathway 英文参考文献.docVIP

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Flagellin-Induced Corneal Antimicrobial Peptide Production and Wound Repair Involve a Novel NF-κB–Independent and EGFR-Dependent Pathway 英文参考文献.doc

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Flagellin-Induced Corneal Antimicrobial Peptide Production and Wound Repair Involve a Novel NF-κB–Independent and EGFR-Dependent Pathway 英文参考文献

Flagellin-InducedCornealAntimicrobialPeptide ProductionandWoundRepairInvolveaNovel NF-kB–IndependentandEGFR-DependentPathway NanGao1,AshokKumar1,JeevanJyot2,Fu-ShinYu1* 1DepartmentsofOphthalmology,Anatomy,andCellBiology,WayneStateUniversitySchoolofMedicine,Detroit,Michigan,UnitedStatesofAmerica,2Departmentof Medicine/InfectiousDiseases,UniversityofFlorida,Gainesville,Florida,UnitedStatesofAmerica Abstract Background:Thebacterialproteinflagellinplaysamajorroleinstimulatingmucosalsurfaceinnateimmuneresponseto bacterialinfectionanduniquelyinducesprofoundcytoprotectionagainstpathogens,chemicals,andradiation.Thisstudy soughttodeterminesignalingpathwaysresponsiblefortheflagellin-inducedinflammatoryandcytoprotectiveeffectson humancornealepithelialcells(HCECs). Methodology/PrincipalFindings:FlagellinpurifiedfromPseudomonasaeruginosa(strainPAK)orlivebacteriawereusedto challenge cultured HCECs. The activation of signaling pathways was assessed with Western blot, and the secretion of cytokine/chemokineandproductionofantimicrobialpeptides(AMPs)weremeasuredwithELISAanddotblot,respectively. Effects of flagellin on wound healing were assessed in cultured porcine corneas. L94A (a site mutation in TLR5 binding region)flagellinandPAKexpressingL94AflagellinwereunabletostimulateNF-kBactivation,butwerepotentineliciting EGFR signaling in a TGF-a–related pathway in HCECs. Concomitant with the lack of NF-kB activation, L94A flagellin was ineffectiveininducingIL-6andIL-8productioninHCECs.Surprisingly,thesecretionoftwoinducibleAMPs,LL-37andhBD2, wasnotaffectedbyL94Amutation.Similartowild-typeflagellin,L94Ainducedepithelialwoundclosureinculturedporcine corneathroughmaintainingEGFR-mediatedsignaling. Conclusions/Significance: Our data suggest that inflammatory response mediated by NF-kB can be uncoupled from epithelialinnatedefensemachinery(i.e.,AMPexpression)andmajorepithelialproliferation/repairpathwaysmediatedby EGFR,andthatflagellinanditsderivativesmayhavebroadtherapeuticapplications