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B in rat pancreatic acinar cells
Department of Physiology, University of Michigan Medical School, Ann Arbor, Michigan 48109-0622
In the cholecystokinin (CCK)
hyperstimulation model of acute pancreatitis, two early intracellular
events, activation of trypsinogen and activation of nuclear factor-
B
(NF-B), are thought to be important in the development of the
disease. In this study, the relationship between these two events was
investigated. NF-B activity was monitored by using a DNA binding
assay and mob-1 chemokine gene expression. Intracellular
trypsin activity was measured by using a fluorogenic substrate.
Protease inhibitors including FUT-175, Pefabloc, and E-64d prevented
CCK stimulation of intracellular trypsinogen and NF-B activation.
Likewise, the NF-B inhibitors pyrrolidine dithiocarbamate and
N-acetyl-L-cysteine inhibited CCK stimulation of
NF-B and intracellular trypsinogen activation. These results
suggested a possible codependency of these two events. However, CCK
stimulated NF-B activation in Chinese hamster ovary-CCKA
cells, which do not express trypsinogen, indicating that trypsin is not
necessary for CCK activation of NF-B. Furthermore,
adenovirus-mediated expression in acinar cells of active p65 subunits
to stimulate NF-B, or of inhibitory B-
molecules to inhibit
NF-B, did not affect either basal or CCK-mediated trypsinogen
activation. Thus trypsinogen and NF-B activation are independent
events stimulated by CCK.
pancreatitis; inflammation; cholecystokinin; nuclear
factor-B
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