Intestinal NADPH oxidase 2 activity increases in a neonatal rat model of necrotizing enterocolitis

Scott R. Welak, Rebecca M. Rentea, Ru Jeng Teng, Nathan Heinzerling, Ben Biesterveld, Jennifer L. Liedel, Kirkwood A. Pritchard, Katherine M. Fredrich, David M. Gourlay

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Necrotizing enterocolitis (NEC) is a complication of prematurity. The etiology is unknown, but is related to enteral feeding, ischemia, infection, and inflammation. Reactive oxygen species production, most notably superoxide, increases in NEC. NADPH oxidase (NOX) generates superoxide, but its activity in NEC remains unknown. We hypothesize that NOX-derived superoxide production increases in NEC. Newborn Sprague-Dawley rats were divided into control, formula-fed, formula/LPS, formula/hypoxia, and NEC (formula, hypoxia, and LPS). Intestinal homogenates were analyzed for NADPH-dependent superoxide production. Changes in superoxide levels on days 0-4 were measured. Inhibitors for nitric oxide synthase (L-NAME) and NOX2 (GP91-ds-tat) were utilized. RT-PCR for eNOS, NOX1, GP91phox expression was performed. Immunofluorescence studies estimated the co-localization of p47phox and GP91phox in control and NEC animals on D1, D2, and D4. NEC pups generated more superoxide than controls on D4, while all other groups were unchanged. NADPH-dependent superoxide production was greater in NEC on days 0, 3, and 4. GP91-ds-tat decreased superoxide production in both groups, with greater inhibition in NEC. L-NAME did not alter superoxide production. Temporally, superoxide production varied minimally in controls. In NEC, superoxide generation was decreased on day 1, but increased on days 3-4. GP91phox expression was higher in NEC on days 2 and 4. NOX1 and eNOS expression were unchanged from controls. GP91phox and p47phox had minimal co-localization in all control samples and NEC samples on D1 and D2, but had increased co-localization on D4. In conclusion, this study proves that experimentally-induced NEC increases small intestinal NOX activity. All components of NEC model are necessary for increased NOX activity. NOX2 is the major source, especially as the disease progresses.

Original languageEnglish (US)
Article numbere115317
JournalPLoS One
Volume9
Issue number12
DOIs
StatePublished - Dec 17 2014
Externally publishedYes

Fingerprint

enterocolitis
Necrotizing Enterocolitis
NADPH Oxidase
Superoxides
Rats
neonates
animal models
superoxide anion
NG-Nitroarginine Methyl Ester
NADP
NAD(P)H oxidase (H2O2-forming)
NADP (coenzyme)
hypoxia
premature birth
Nitric Oxide Synthase
feed formulation
enteral feeding
Enteral Nutrition
Reactive Oxygen Species
Animals

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Welak, S. R., Rentea, R. M., Teng, R. J., Heinzerling, N., Biesterveld, B., Liedel, J. L., ... Gourlay, D. M. (2014). Intestinal NADPH oxidase 2 activity increases in a neonatal rat model of necrotizing enterocolitis. PLoS One, 9(12), [e115317]. https://doi.org/10.1371/journal.pone.0115317

Intestinal NADPH oxidase 2 activity increases in a neonatal rat model of necrotizing enterocolitis. / Welak, Scott R.; Rentea, Rebecca M.; Teng, Ru Jeng; Heinzerling, Nathan; Biesterveld, Ben; Liedel, Jennifer L.; Pritchard, Kirkwood A.; Fredrich, Katherine M.; Gourlay, David M.

In: PLoS One, Vol. 9, No. 12, e115317, 17.12.2014.

Research output: Contribution to journalArticle

Welak, SR, Rentea, RM, Teng, RJ, Heinzerling, N, Biesterveld, B, Liedel, JL, Pritchard, KA, Fredrich, KM & Gourlay, DM 2014, 'Intestinal NADPH oxidase 2 activity increases in a neonatal rat model of necrotizing enterocolitis', PLoS One, vol. 9, no. 12, e115317. https://doi.org/10.1371/journal.pone.0115317
Welak SR, Rentea RM, Teng RJ, Heinzerling N, Biesterveld B, Liedel JL et al. Intestinal NADPH oxidase 2 activity increases in a neonatal rat model of necrotizing enterocolitis. PLoS One. 2014 Dec 17;9(12). e115317. https://doi.org/10.1371/journal.pone.0115317
Welak, Scott R. ; Rentea, Rebecca M. ; Teng, Ru Jeng ; Heinzerling, Nathan ; Biesterveld, Ben ; Liedel, Jennifer L. ; Pritchard, Kirkwood A. ; Fredrich, Katherine M. ; Gourlay, David M. / Intestinal NADPH oxidase 2 activity increases in a neonatal rat model of necrotizing enterocolitis. In: PLoS One. 2014 ; Vol. 9, No. 12.
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