Dominant negative p38 mitogen-activated protein kinase expression inhibits NF-kappaB activation in AR42J cells.
BACKGROUND
The role of the p38 mitogen-activated protein (MAP) kinase in acute pancreatitis pathogenesis is controversial. We hypothesize that p38 plays a role in regulating NF-kappaB activation in exocrine pancreatic cells.


METHODS
AR42J cells incorporating an NF-kappaB-responsive luciferase reporter, with and without adenoviral transduction of DNp38, were stimulated with cholecystokinin (CCK) or tumor necrosis factor-alpha (TNF-alpha) prior to measuring NF-kappaB activation.


RESULTS
CCK- or TNF-alpha-stimulated NF-kappaB-dependent gene transcription (luciferase assay) was substantially subdued by DNp38 expression. These findings were confirmed by electrophoretic mobility shift assay. Nuclear translocation of the p65 NF-kappaB subunit following agonist stimulation was evident (supershift). Characterization studies showed excellent adenoviral infection efficiency and cell viability in our AR42J cell model. Agonist-stimulated dose- and time-dependent p38 activation, with inhibition by DNp38 expression, was also confirmed.


CONCLUSIONS
The p38 MAP kinase regulates NF-kappaB pathway activation in exocrine pancreatic cells, and thus potentially plays a role in the mechanism of acute pancreatitis pathogenesis..
Twait
Erik
E
Surgical Services, Iowa City Veterans Affairs Medical Center, and Department of Surgery, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa, USA.
Williard
Deborah E
DE
Samuel
Isaac
I
D000818:Animals; D002460:Cell Line; D002766:Cholecystokinin / Q000494:pharmacology; D004789:Enzyme Activation; D005799:Genes, Dominant; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism*; D010179:Pancreas / Q000378:metabolism*; D010195:Pancreatitis / Q000209:etiology; D051381:Rats; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000502:physiology*
D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D002766:Cholecystokinin; D048051:p38 Mitogen-Activated Protein Kinases
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