P38 MAP kinase mediates endotoxin-induced expression of cyclooxygenase-2 in enterocytes

Anatoly Grishin, Jin Wang, David Hackam, Faisal Qureshi, Jeffrey Upperman, Ruben Zamora, Henri R. Ford

Research output: Contribution to journalArticle

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Abstract

Background Necrotizing enterocolitis (NEC) occurs only after bacterial colonization of the intestine, suggesting that bacterial products, including lipopolysaccharide (endotoxin,) interact with enterocytes in the pathogenesis of this disease. Inflammatory molecules such as cyclooxygenase-2 (COX-2) are important mediators of the septic response leading to NEC. We therefore hypothesized that endotoxin activates production of COX-2 in enterocytes and explored the relative contributions of known mitogen-activated protein kinases (MAPK) pathways in this process. Methods IEC-6 enterocytes were treated with 5 μg/mL endotoxin, or various stresses, or media alone, and COX-2 protein levels were assayed by immunoblots with anti-COX-2 antibodies. Activation of MAPK was examined by immunoblots with phospho-MAPK antibodies. MAPK activity was blocked by treatment with pharmacologic inhibitors or transfection with dominant-negative MAPK constructs. Results Endotoxin treatment caused increased expression of the COX-2 protein 24 hours after treatment. This was preceded by rapid and transient activation of the 3 major MAPKs: extracellular-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. SB203580, a specific inhibitor of p38, but not U0126 (ERK inhibitor) or SP600125 (JNK inhibitor), blocked endotoxin-induced accumulation of COX-2 protein. This response was also blocked by expression of dominant-negative p38 but not by the dominant-negative ERK construct. Genotoxic stress that activated p38 but not ERK was an effective inducer of COX-2, whereas stresses that activated both p38 and ERK were not effective. ERK inhibition by U1026 enhanced endotoxin-induced production of COX-2, consistent with negative regulation of COX-2 by ERK. These data point to p38 as the MAPK that mediates endotoxin-induced production of COX-2 in enterocytes. Conclusions Endotoxin may be capable of inducing the production of COX-2 in enterocytes via the p38 MAPK pathway, which may be relevant to the development of NEC.

Original languageEnglish (US)
Pages (from-to)329-335
Number of pages7
JournalSurgery
Volume136
Issue number2
DOIs
StatePublished - Aug 1 2004

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Enterocytes
p38 Mitogen-Activated Protein Kinases
Cyclooxygenase 2
Endotoxins
Phosphotransferases
Mitogen-Activated Protein Kinases
Necrotizing Enterocolitis
Proteins
Antibodies
JNK Mitogen-Activated Protein Kinases
DNA Damage
Intestines
Transfection
Lipopolysaccharides

ASJC Scopus subject areas

  • Surgery

Cite this

Grishin, A., Wang, J., Hackam, D., Qureshi, F., Upperman, J., Zamora, R., & Ford, H. R. (2004). P38 MAP kinase mediates endotoxin-induced expression of cyclooxygenase-2 in enterocytes. Surgery, 136(2), 329-335. https://doi.org/10.1016/j.surg.2004.05.008

P38 MAP kinase mediates endotoxin-induced expression of cyclooxygenase-2 in enterocytes. / Grishin, Anatoly; Wang, Jin; Hackam, David; Qureshi, Faisal; Upperman, Jeffrey; Zamora, Ruben; Ford, Henri R.

In: Surgery, Vol. 136, No. 2, 01.08.2004, p. 329-335.

Research output: Contribution to journalArticle

Grishin, A, Wang, J, Hackam, D, Qureshi, F, Upperman, J, Zamora, R & Ford, HR 2004, 'P38 MAP kinase mediates endotoxin-induced expression of cyclooxygenase-2 in enterocytes', Surgery, vol. 136, no. 2, pp. 329-335. https://doi.org/10.1016/j.surg.2004.05.008
Grishin, Anatoly ; Wang, Jin ; Hackam, David ; Qureshi, Faisal ; Upperman, Jeffrey ; Zamora, Ruben ; Ford, Henri R. / P38 MAP kinase mediates endotoxin-induced expression of cyclooxygenase-2 in enterocytes. In: Surgery. 2004 ; Vol. 136, No. 2. pp. 329-335.
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abstract = "Background Necrotizing enterocolitis (NEC) occurs only after bacterial colonization of the intestine, suggesting that bacterial products, including lipopolysaccharide (endotoxin,) interact with enterocytes in the pathogenesis of this disease. Inflammatory molecules such as cyclooxygenase-2 (COX-2) are important mediators of the septic response leading to NEC. We therefore hypothesized that endotoxin activates production of COX-2 in enterocytes and explored the relative contributions of known mitogen-activated protein kinases (MAPK) pathways in this process. Methods IEC-6 enterocytes were treated with 5 μg/mL endotoxin, or various stresses, or media alone, and COX-2 protein levels were assayed by immunoblots with anti-COX-2 antibodies. Activation of MAPK was examined by immunoblots with phospho-MAPK antibodies. MAPK activity was blocked by treatment with pharmacologic inhibitors or transfection with dominant-negative MAPK constructs. Results Endotoxin treatment caused increased expression of the COX-2 protein 24 hours after treatment. This was preceded by rapid and transient activation of the 3 major MAPKs: extracellular-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. SB203580, a specific inhibitor of p38, but not U0126 (ERK inhibitor) or SP600125 (JNK inhibitor), blocked endotoxin-induced accumulation of COX-2 protein. This response was also blocked by expression of dominant-negative p38 but not by the dominant-negative ERK construct. Genotoxic stress that activated p38 but not ERK was an effective inducer of COX-2, whereas stresses that activated both p38 and ERK were not effective. ERK inhibition by U1026 enhanced endotoxin-induced production of COX-2, consistent with negative regulation of COX-2 by ERK. These data point to p38 as the MAPK that mediates endotoxin-induced production of COX-2 in enterocytes. Conclusions Endotoxin may be capable of inducing the production of COX-2 in enterocytes via the p38 MAPK pathway, which may be relevant to the development of NEC.",
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AU - Grishin, Anatoly

AU - Wang, Jin

AU - Hackam, David

AU - Qureshi, Faisal

AU - Upperman, Jeffrey

AU - Zamora, Ruben

AU - Ford, Henri R.

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N2 - Background Necrotizing enterocolitis (NEC) occurs only after bacterial colonization of the intestine, suggesting that bacterial products, including lipopolysaccharide (endotoxin,) interact with enterocytes in the pathogenesis of this disease. Inflammatory molecules such as cyclooxygenase-2 (COX-2) are important mediators of the septic response leading to NEC. We therefore hypothesized that endotoxin activates production of COX-2 in enterocytes and explored the relative contributions of known mitogen-activated protein kinases (MAPK) pathways in this process. Methods IEC-6 enterocytes were treated with 5 μg/mL endotoxin, or various stresses, or media alone, and COX-2 protein levels were assayed by immunoblots with anti-COX-2 antibodies. Activation of MAPK was examined by immunoblots with phospho-MAPK antibodies. MAPK activity was blocked by treatment with pharmacologic inhibitors or transfection with dominant-negative MAPK constructs. Results Endotoxin treatment caused increased expression of the COX-2 protein 24 hours after treatment. This was preceded by rapid and transient activation of the 3 major MAPKs: extracellular-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. SB203580, a specific inhibitor of p38, but not U0126 (ERK inhibitor) or SP600125 (JNK inhibitor), blocked endotoxin-induced accumulation of COX-2 protein. This response was also blocked by expression of dominant-negative p38 but not by the dominant-negative ERK construct. Genotoxic stress that activated p38 but not ERK was an effective inducer of COX-2, whereas stresses that activated both p38 and ERK were not effective. ERK inhibition by U1026 enhanced endotoxin-induced production of COX-2, consistent with negative regulation of COX-2 by ERK. These data point to p38 as the MAPK that mediates endotoxin-induced production of COX-2 in enterocytes. Conclusions Endotoxin may be capable of inducing the production of COX-2 in enterocytes via the p38 MAPK pathway, which may be relevant to the development of NEC.

AB - Background Necrotizing enterocolitis (NEC) occurs only after bacterial colonization of the intestine, suggesting that bacterial products, including lipopolysaccharide (endotoxin,) interact with enterocytes in the pathogenesis of this disease. Inflammatory molecules such as cyclooxygenase-2 (COX-2) are important mediators of the septic response leading to NEC. We therefore hypothesized that endotoxin activates production of COX-2 in enterocytes and explored the relative contributions of known mitogen-activated protein kinases (MAPK) pathways in this process. Methods IEC-6 enterocytes were treated with 5 μg/mL endotoxin, or various stresses, or media alone, and COX-2 protein levels were assayed by immunoblots with anti-COX-2 antibodies. Activation of MAPK was examined by immunoblots with phospho-MAPK antibodies. MAPK activity was blocked by treatment with pharmacologic inhibitors or transfection with dominant-negative MAPK constructs. Results Endotoxin treatment caused increased expression of the COX-2 protein 24 hours after treatment. This was preceded by rapid and transient activation of the 3 major MAPKs: extracellular-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. SB203580, a specific inhibitor of p38, but not U0126 (ERK inhibitor) or SP600125 (JNK inhibitor), blocked endotoxin-induced accumulation of COX-2 protein. This response was also blocked by expression of dominant-negative p38 but not by the dominant-negative ERK construct. Genotoxic stress that activated p38 but not ERK was an effective inducer of COX-2, whereas stresses that activated both p38 and ERK were not effective. ERK inhibition by U1026 enhanced endotoxin-induced production of COX-2, consistent with negative regulation of COX-2 by ERK. These data point to p38 as the MAPK that mediates endotoxin-induced production of COX-2 in enterocytes. Conclusions Endotoxin may be capable of inducing the production of COX-2 in enterocytes via the p38 MAPK pathway, which may be relevant to the development of NEC.

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