Expression of TLR-2, TLR-4, NOD2 and pNF-κB in a Neonatal Rat Model of Necrotizing Enterocolitis 英文参考文献.docVIP

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Expression of TLR-2, TLR-4, NOD2 and pNF-κB in a Neonatal Rat Model of Necrotizing Enterocolitis 英文参考文献.doc

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Expression of TLR-2, TLR-4, NOD2 and pNF-κB in a Neonatal Rat Model of Necrotizing Enterocolitis 英文参考文献

ExpressionofTLR-2,TLR-4,NOD2andpNF-k Bin aNeonatalRatModelofNecrotizingEnterocolitis AurelieLeMandatSchultz1,2.,ArnaudBonnard1,2.,Fre′de′rickBarreau5,6,YvesAigrain1,2,3,CoraliePierre-Louis2,4,DominiqueBerrebi2,4.*, MichelPeuchmaur2,4. 1AssistancePublique-Ho?pitauxdeParis(AP-HP)Ho?pitalR.Debre′,ServicedeChirurgiePe′diatrique, Paris,France,2Universite′ Paris7,EA3102,Paris, France,3EcoledeChirurgie,AssistancePublique-Ho?pitauxdeParis(AP-HP),Paris,France,4AssistancePublique-Ho?pitauxdeParis(AP-HP)Ho?pital RobertDebre′,Serviced’Anatomieet deCytologiePathologiques, Paris,France,5INSERM,U843,Ho?pitalRobertDebre′,Paris,France,6UMR-S843, Universite′ ParisDiderot,Paris,France Background. The etiology of necrotizing enterocolitis (NEC) results from a combination of several risk factors that act synergisticallyandoccursinthesamecircumstancesasthosewhichleadtoinnateimmunityactivation.Patternrecognition moleculescouldbeanimportantplayerintheinitiationofanexaggeratedinflammatoryresponseleadingtointestinalinjury in NEC. Methodology/Principal Findings. We specifically evaluated intestinal epithelial cell (IEC) expression of Toll-like receptor2(TLR-2),TLR-4,NOD2andphosphorylatedNF-kB(pNF-kB)aftermucosalinjuryinaratmodelofNECinducedby prematurity, systemic hypoxia, and a rich protein formula. In the control group (group1), neonatal rats were full-term and breast-fed; in the experimental groups, rat pups were preterm at day 21 of gestation and rat-milk fed (group 2) or hand- gavagedwithaproteinrichformulaafterahypoxia–reoxygenationprocedure(group3).Morphologicalmucosalchangesin the small bowel were scored on hematoxylin- and eosin-stained sections. Immunohistochemistry was performed on frozen tissuesectionsusingantiTLR-2andactivepNF-kBp65antibodies.Real-timeRT-PCRwasperformedtoassessmRNAexpression of NOD2, TLR-2 and TLR-4. Proliferation and apoptosis were studied in paraffin sections using anti Ki-67 and caspase-3 antibodies,respectively.Thecombinationofimmaturity,proteinri