title: PKC-delta and -epsilon regulate NF-kappaB activation induced by cholecystokinin and TNF-alpha in pancreatic acinar cells.
abstract: Although NF-kappaB plays an important role in pancreatitis, mechanisms underlying its activation remain unclear. We investigated the signaling pathways mediating NF-kappaB activation in pancreatic acinar cells induced by high-dose cholecystokinin-8 (CCK-8), which causes pancreatitis in rodent models, and TNF-alpha, which contributes to inflammatory responses of pancreatitis, especially the role of PKC isoforms. We determined subcellular distribution and kinase activities of PKC isoforms and NF-kappaB activation in dispersed rat pancreatic acini. We applied isoform-specific, cell-permeable peptide inhibitors to assess the role of individual PKC isoforms in NF-kappaB activation. Both CCK-8 and TNF-alpha activated the novel isoforms PKC-delta and -epsilon and the atypical isoform PKC-zeta but not the conventional isoform PKC-alpha. Inhibition of the novel PKC isoforms but not the conventional or the atypical isoform resulted in the prevention of NF-kappaB activation induced by CCK-8 and TNF-alpha. NF-kappaB activation by CCK-8 and TNF-alpha required translocation but not tyrosine phosphorylation of PKC-delta. Activation of PKC-delta, PKC-epsilon, and NF-kappaB with CCK-8 involved both phosphatidylinositol-specific PLC and phosphatidylcholine (PC)-specific PLC, whereas with TNF-alpha they only required PC-specific PLC for activation. Results indicate that CCK-8 and TNF-alpha initiate NF-kappaB activation by different PLC pathways that converge at the novel PKCs (delta and epsilon) to mediate NF-kappaB activation in pancreatic acinar cells. These findings suggest a key role for the novel PKCs in pancreatitis.
authors.affiliation: Research Center for Alcoholic Liver and Pancreatic Diseases, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, California 90073, USA.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Satoh
authors.forename: Anna S
authors.initials: AS
authors.lastname: Gukovskaya
authors.forename: Jose M
authors.initials: JM
authors.lastname: Nieto
authors.forename: Jason H
authors.initials: JH
authors.lastname: Cheng
authors.forename: Ilya
authors.initials: I
authors.lastname: Gukovsky
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Reeve
authors.forename: Tooru
authors.initials: T
authors.lastname: Shimosegawa
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Pandol
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D002766:Cholecystokinin / Q000494:pharmacology*; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D024202:Electrophoretic Mobility Shift Assay; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D016328:NF-kappa B / Q000502:physiology*; D010179:Pancreas / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000502:physiology; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism; D019061:src-Family Kinases / Q000037:antagonists & inhibitors
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D002766:Cholecystokinin; C497260:Prkcd protein, rat; C497257:Prkce protein, rat; D019061:src-Family Kinases; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D010738:Type C Phospholipases