title: The roles of bacteria and TLR4 in rat and murine models of necrotizing enterocolitis.
abstract: Bacteria are thought to contribute to the pathogenesis of necrotizing enterocolitis (NEC), but it is unknown whether their interaction with the epithelium can participate in the initiation of mucosal injury or they can act only following translocation across a damaged intestinal barrier. Our aims were to determine whether bacteria and intestinal epithelial TLR4 play roles in a well-established neonatal rat model and a novel neonatal murine model of NEC. Neonatal rats, C57BL/6J, C3HeB/FeJ (TLR4 wild type), and C3H/HeJ (TLR4 mutant) mice were delivered by Cesarean section and were subjected to formula feeding and cold asphyxia stress or were delivered naturally and were mother-fed. NEC incidence was evaluated by histological scoring, and gene expression was quantified using quantitative real-time PCR from cDNA generated from intestinal total RNA or from RNA obtained by laser capture microdissection. Spontaneous feeding catheter colonization or supplementation of cultured bacterial isolates to formula increased the incidence of experimental NEC. During the first 72 h of life, i.e., the time frame of NEC development in this model, intestinal TLR4 mRNA gradually decreases in mother-fed but increases in formula feeding and cold asphyxia stress, correlating with induced inducible NO synthase. TLR4, inducible NO synthase, and inflammatory cytokine induction occurred in the intestinal epithelium but not in the submucosa. NEC incidence was diminished in C3H/HeJ mice, compared with C3HeB/FeJ mice. In summary, bacteria and TLR4 play significant roles in experimental NEC, likely via an interaction of intraluminal bacteria and aberrantly overexpressed TLR4 in enterocytes.
authors.affiliation: Department of Pediatrics, Northwestern University Feinberg School of Medicine, Chicago, IL 60614, USA.
authors.forename: Tamas
authors.initials: T
authors.lastname: Jilling
authors.forename: Dyan
authors.initials: D
authors.lastname: Simon
authors.forename: Jing
authors.initials: J
authors.lastname: Lu
authors.forename: Fan Jing
authors.initials: FJ
authors.lastname: Meng
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.forename: Robert
authors.initials: R
authors.lastname: Schy
authors.forename: Richard B
authors.initials: RB
authors.lastname: Thomson
authors.forename: Antoine
authors.initials: A
authors.lastname: Soliman
authors.forename: Moshe
authors.initials: M
authors.lastname: Arditi
authors.forename: Michael S
authors.initials: MS
authors.lastname: Caplan
mesh_terms: D000821:Animal Feed; D000818:Animals; D001237:Asphyxia / Q000378:metabolism / Q000473:pathology; D019743:Chemokines, CXC / Q000378:metabolism; D002824:Chorionic Villi / Q000378:metabolism; D004195:Disease Models, Animal; D020345:Enterocolitis, Necrotizing / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology* / Q000473:pathology; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D051379:Mice; D009154:Mutation / Q000235:genetics; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D019743:Chemokines, CXC; D012333:RNA, Messenger; C493487:Tlr4 protein, mouse; C495347:Tlr4 protein, rat; D051197:Toll-Like Receptor 4; D052247:Nitric Oxide Synthase Type II