title: Carbon monoxide protects against the development of experimental necrotizing enterocolitis.
abstract: Necrotizing enterocolitis (NEC) is a disease of neonates that is increasing in incidence and often results in significant morbidity and mortality. Carbon monoxide (CO), a byproduct of the catabolism of heme, is known to have anti-inflammatory and antiapoptotic properties. In this study, we aimed to demonstrate that inhaled CO protects against the development of intestinal inflammation in a model of experimental NEC as well as decreases enterocyte cell death in vitro. Additionally, we also aimed to demonstrate that CO decreases enterocyte production of inducible nitric oxide synthase (iNOS) and nitric oxide (NO). Neonatal rats were exposed to intermittent hypoxia exposure and formula feeding to induce experimental NEC. Animals randomized to CO treatment were put in an environment containing 0.025% CO for 1 h/day on days 1-3 of life. All animals were killed on day 4 of life. In vitro experiments were performed with IEC-6 cells, a rat enterocyte cell line. Cells were examined for viability, iNOS production, and elaboration of NO. We found that CO diminished levels of serum inflammatory cytokines and nitrites, protected against intestinal inflammation, and decreased ileal iNOS production and protein nitration in a model of experimental NEC. In vitro, CO decreased cytokine- or hypoxia/endotoxin-induced iNOS and NO production. CO also abrogated TNF-alpha- and actinomycin D-induced apoptosis or hypoxia/endotoxin-induced cell death. In conclusion, 1 h of daily low-dose inhaled CO protected against the development of intestinal inflammation in a model of experimental NEC. iNOS and NO production were decreased by CO both in vivo and in vitro. CO may prove to be a useful clinical adjunct in the treatment of NEC.
authors.affiliation: Dept. of Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. zuckerbraunbs@upmc.edu
authors.forename: Brian S
authors.initials: BS
authors.lastname: Zuckerbraun
authors.forename: Leo E
authors.initials: LE
authors.lastname: Otterbein
authors.forename: Patricia
authors.initials: P
authors.lastname: Boyle
authors.forename: Ronald
authors.initials: R
authors.lastname: Jaffe
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Upperman
authors.forename: Ruben
authors.initials: R
authors.lastname: Zamora
authors.forename: Henri R
authors.initials: HR
authors.lastname: Ford
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D002248:Carbon Monoxide / Q000008:administration & dosage / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002470:Cell Survival; D004195:Disease Models, Animal; D020345:Enterocolitis, Necrotizing / Q000503:physiopathology* / Q000517:prevention & control*; D020895:Enterocytes; D000860:Hypoxia; D007249:Inflammation; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D052247:Nitric Oxide Synthase Type II; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002248:Carbon Monoxide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat