title: Stimulatory effects of vanadate on amylase release from isolated rat pancreatic acini.
abstract: The effects of vanadate on exocrine pancreatic function were examined in isolated rat pancreatic acini. Vanadate caused a concentration-dependent stimulation of amylase release above a concentration of 1 mM. Co-incubation of vanadate with vasoactive intestinal polypeptide, 8-bromoadenosine 3':5'-cyclic monophosphate, and the Ca2+ ionophore A23187 produced a synergistic pattern of amylase release, whereas co-incubation with cholecystokinin octapeptide (CCK-8), carbamylcholine, and 12-O-tetradecanoylphorbol 13-acetate produced an additive effect. Vanadate alone had no influence on acinar cyclic AMP content, Ca2+ efflux, or intracellular Ca2+ concentration. However, preincubation with vanadate prevented the plateau phase of CCK-8-induced Ca2+ transient increase from returning to baseline. Moreover, depletion of the intracellular Ca2+ pool by pretreatment of acini with CCK-8 in Ca2+-free medium (plus ethyleneglycol bis[beta-aminoethylether]-N,N'-tetraacetic acid) had no effect on subsequent stimulation by vanadate, although it abolished the response to both CCK-8 and carbamylcholine stimulation. The protein kinase C (PKC) inhibitors staurosporine and calphostin C significantly inhibited vanadate-stimulated amylase release, whereas the protein tyrosine kinase inhibitor genistein had no inhibitory effect. Moreover, vanadate caused a significant translocation of PKC from cytosol to membrane fraction in pancreatic acinar cells. This translocation was inhibited significantly by staurosporine and calphostin C but not by genistein. These results suggest that vanadate acts directly on pancreatic acini and stimulates amylase release by activating PKC without an effect on Ca2+ mobilization, cyclic AMP, or protein tyrosine kinase.
authors.affiliation: Third Department of Internal Medicine, University of Occupational and Environmental Health, Japan, School of Medicine, Kitakyushu.
authors.forename: Y
authors.initials: Y
authors.lastname: Hirohata
authors.forename: Y
authors.initials: Y
authors.lastname: Ogami
authors.forename: T
authors.initials: T
authors.lastname: Akiyama
authors.forename: I
authors.initials: I
authors.lastname: Shibuya
authors.forename: M
authors.initials: M
authors.lastname: Otsuki
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000681:Amylases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D004347:Drug Interactions; D004789:Enzyme Activation; D017136:Ion Transport; D008297:Male; D010179:Pancreas / Q000187:drug effects* / Q000201:enzymology; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D012844:Sincalide / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology*; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
chemical_list: D015124:8-Bromo Cyclic Adenosine Monophosphate; D014660:Vasoactive Intestinal Peptide; D014638:Vanadates; D002217:Carbachol; D011493:Protein Kinase C; D000681:Amylases; D012844:Sincalide; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium