Bacteria Regulate Intestinal Epithelial Cell Differentiation Factors Both In Vitro and In Vivo.docVIP

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2017-05-07 发布于上海
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Bacteria Regulate Intestinal Epithelial Cell Differentiation Factors Both In Vitro and In Vivo.doc

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Bacteria Regulate Intestinal Epithelial Cell Differentiation Factors Both In Vitro and In Vivo

BacteriaRegulateIntestinalEpithelialCellDifferentiation FactorsBothInVitroandInVivo SvetlanaBecker1,2,TobiasA.Oelschlaeger3,AndyWullaert4,ManolisPasparakis4,5,JanWehkamp1,2,6 , EduardF.Stange6,MichaelGersemann1,2,6* 1Dr.MargareteFischer-BoschInstituteforClinicalPharmacology,Stuttgart,Germany,2UniversityofTu¨bingen,Tu¨bingen,Germany,3InstituteforMolecularBiologyof Infection,UniversityofWu¨rzburg,Wu¨rzburg,Germany,4Institute forGenetics,UniversityofCologne,Cologne,Germany, 5EMBLMouseBiologyUnit,Monterotondo, Italy,6DepartmentofInternalMedicineI,RobertBoschHospital,Stuttgart,Germany Abstract Background: The human colon harbours a plethora of bacteria known to broadly impact on mucosal metabolism and functionandthoughttobeinvolvedininflammatoryboweldiseasepathogenesisandcoloncancerdevelopment.Inthis report, we investigated the effect of colonic bacteria on epithelial cell differentiation factors invitro and invivo. As key transcriptionfactorswefocusedonHes1,knowntodirecttowardsanabsorptivecellfate,Hath1andKLF4,whichgovern gobletcell. Methods:ExpressionofthetranscriptionfactorsHes1,Hath1andKLF4,themucinsMuc1andMuc2andthedefensinHBD2 weremeasuredbyreal-timePCRinLS174Tcellsfollowingincubationwithseveralheat-inactivatedE.colistrains,including theprobioticE.coliNissle1917+/2flagellin,LactobacilliandBifidobacteria.ForproteindetectionWesternblotexperiments andchamber-slideimmunostainingwereperformed.Finally,mRNAandproteinexpressionofthesefactorswasevaluatedin thecolonofgermfreevs.specificpathogenfreevs.conventionalizedmiceandcolonicgobletcellswerecounted. Results:ExpressionofHes1andHath1,andtoaminordegreealsoofKLF4,wasreducedbyE.coliK-12andE.coliNissle 1917.Incontrast,Muc1andHBD2expressionweresignificantlyenhanced,independentoftheNotchsignallingpathway. ProbioticE.coliNissle1917regulatedHes1,Hath1,Muc1andHBD2throughflagellin.Invivoexperimentsconfirmedthe observedinvitroeffectsofbacteriabyadiminishedcolonicexpressionofHath1andKLF4inspecificpathogenfreeand conventionalize