Lipopolysaccharide induces cyclooxygenase-2 in intestinal epithelium via a noncanonical p38 MAPK pathway

Anatoly V. Grishin, Jin Wang, Douglas A. Potoka, David J. Hackam, Jeffrey S. Upperman, Patricia Boyle, Ruben Zamora, Henri Ford

Research output: Contribution to journalArticle

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Abstract

Necrotizing enterocolitis (NEC), a severe intestinal inflammation in neonates, occurs following bacterial colonization of the gut. LPS-induced production of inflammatory factors in immature enterocytes may be a factor in NEC. Previously, we described LPS-induced p38 MAPK-dependent expression of cyclooxygenase-2 (COX-2) in rat IEC-6 cells. In this study, we examine COX-2 expression in newborn rat intestinal epithelium and further characterize the mechanisms of COX-2 regulation in enterocytes. Induction of NEC by formula feeding/hypoxia increased phospho-p38 and COX-2 levels in the intestinal mucosa. Celecoxib, a selective COX-2 inhibitor, exacerbated the disease, suggesting a protective role for COX-2. COX-2 was induced in the intestinal epithelium by LPS in vivo and ex vivo. The latter response was attenuated by the p38 inhibitor SB202190, but not by inhibitors of ERK, JNK, or NF-κB. In IEC-6 enterocytes, COX-2 was induced by the expression of MAPK kinase 3 EE (MKK3EE), a constitutive activator of p38, but not of activators of ERK or JNK pathways. However, neither MKK3/6 nor MKK4, the known p38 upstream kinases, were activated by LPS. Dominant-negative MKK3 or MKK4 or SB202190 failed to prevent LPS-induced, p38-activating phosphorylation, ruling out important roles of these kinases or p38 autophosphorylation. LPS increased COX-2 and activating phosphorylation of p38 with similar dose-response. Blockade of LPS-induced expression of COX-2-luciferase reporter and destabilization of COX-2 message by SB202190 indicate that p38 regulates COX-2 at transcription and mRNA stability levels. Our data indicate that p38-mediated expression of COX-2 proceeds through a novel upstream pathway and support the role of the neonate's enterocytes as bacterial sensors.

Original languageEnglish (US)
Pages (from-to)580-588
Number of pages9
JournalJournal of Immunology
Volume176
Issue number1
DOIs
StatePublished - Jan 1 2006
Externally publishedYes

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p38 Mitogen-Activated Protein Kinases
Cyclooxygenase 2
Intestinal Mucosa
Lipopolysaccharides
Enterocytes
Necrotizing Enterocolitis
MAP Kinase Signaling System
Celecoxib
Phosphotransferases
Phosphorylation
Cyclooxygenase 2 Inhibitors
Mitogen-Activated Protein Kinase Kinases
RNA Stability
Luciferases
Inflammation

ASJC Scopus subject areas

  • Immunology

Cite this

Lipopolysaccharide induces cyclooxygenase-2 in intestinal epithelium via a noncanonical p38 MAPK pathway. / Grishin, Anatoly V.; Wang, Jin; Potoka, Douglas A.; Hackam, David J.; Upperman, Jeffrey S.; Boyle, Patricia; Zamora, Ruben; Ford, Henri.

In: Journal of Immunology, Vol. 176, No. 1, 01.01.2006, p. 580-588.

Research output: Contribution to journalArticle

Grishin, AV, Wang, J, Potoka, DA, Hackam, DJ, Upperman, JS, Boyle, P, Zamora, R & Ford, H 2006, 'Lipopolysaccharide induces cyclooxygenase-2 in intestinal epithelium via a noncanonical p38 MAPK pathway', Journal of Immunology, vol. 176, no. 1, pp. 580-588. https://doi.org/10.4049/jimmunol.176.1.580
Grishin, Anatoly V. ; Wang, Jin ; Potoka, Douglas A. ; Hackam, David J. ; Upperman, Jeffrey S. ; Boyle, Patricia ; Zamora, Ruben ; Ford, Henri. / Lipopolysaccharide induces cyclooxygenase-2 in intestinal epithelium via a noncanonical p38 MAPK pathway. In: Journal of Immunology. 2006 ; Vol. 176, No. 1. pp. 580-588.
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AB - Necrotizing enterocolitis (NEC), a severe intestinal inflammation in neonates, occurs following bacterial colonization of the gut. LPS-induced production of inflammatory factors in immature enterocytes may be a factor in NEC. Previously, we described LPS-induced p38 MAPK-dependent expression of cyclooxygenase-2 (COX-2) in rat IEC-6 cells. In this study, we examine COX-2 expression in newborn rat intestinal epithelium and further characterize the mechanisms of COX-2 regulation in enterocytes. Induction of NEC by formula feeding/hypoxia increased phospho-p38 and COX-2 levels in the intestinal mucosa. Celecoxib, a selective COX-2 inhibitor, exacerbated the disease, suggesting a protective role for COX-2. COX-2 was induced in the intestinal epithelium by LPS in vivo and ex vivo. The latter response was attenuated by the p38 inhibitor SB202190, but not by inhibitors of ERK, JNK, or NF-κB. In IEC-6 enterocytes, COX-2 was induced by the expression of MAPK kinase 3 EE (MKK3EE), a constitutive activator of p38, but not of activators of ERK or JNK pathways. However, neither MKK3/6 nor MKK4, the known p38 upstream kinases, were activated by LPS. Dominant-negative MKK3 or MKK4 or SB202190 failed to prevent LPS-induced, p38-activating phosphorylation, ruling out important roles of these kinases or p38 autophosphorylation. LPS increased COX-2 and activating phosphorylation of p38 with similar dose-response. Blockade of LPS-induced expression of COX-2-luciferase reporter and destabilization of COX-2 message by SB202190 indicate that p38 regulates COX-2 at transcription and mRNA stability levels. Our data indicate that p38-mediated expression of COX-2 proceeds through a novel upstream pathway and support the role of the neonate's enterocytes as bacterial sensors.

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