Liver, Pancreas, and Biliary TractSerine protease inhibitor causes F-actin redistribution and inhibition of calcium-mediated secretion in pancreatic acini*,**
Section snippets
Materials and methods
Male Wistar rats (90–120 g) were obtained from Charles River Laboratories (Wilmington, MA). Cerulein was purchased from Research Plus (Bayonne, NJ), collagenase from Worthington Biochemicals (Freehold, NJ), the trypsin substrate Boc-Glu-Ala-Arg-4-methylcoumaryl-7-amide (MCA) from Peptides International (Louisville, KY), and both Fura 2/AM and rhodamine phalloidin from Molecular Probes (Eugene, OR). The serine protease substrate tosyl-glycyl-prylyl-arginine-4-nitralide acetate (Chromozym TH) was
Cell-permeant serine protease inhibitors cause inhibition of Ca2+-mediated but not camp-mediated amylase secretion from intact acini
Cerulein-stimulated amylase secretion from freshly prepared acini is inhibited, in a concentration-dependent manner, by AEBSF, as well as the other cell-permeant serine protease inhibitors TPCK and TLCK (Figure 1).
Discussion
Pancreatic acinar cell stimulus-secretion coupling involves 2 parallel intracellular second messenger systems, 1 that results in an increase in [Ca2+]I and activation of protein kinase C, and another that results in generation of cAMP and activation of protein kinase A. In this article, we have shown that exposure to a cell-permeant serine protease inhibitor selectively interferes with secretion mediated via the Ca2+ pathway but does not alter secretion mediated via the cAMP pathway.
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2023, Vitamins and HormonesLiquid crystal-enabled protease inhibition assays developed in a millifluidic device
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Fatty acid ethyl esters are less toxic than their parent fatty acids generated during acute pancreatitis
2016, American Journal of PathologyCitation Excerpt :All experiments were approved by the Institutional Animal Care and Use Committee at the University of Pittsburgh and the Mayo Clinic (Scottsdale, AZ). Mice pancreatic acini were harvested, as described previously.30,31 Fresh acini were used for all assays.
Acute lipotoxicity regulates severity of biliary acute pancreatitis without affecting its initiation
2014, American Journal of PathologyCitation Excerpt :All experiments were approved by the Institutional Animal Care and Use Committee of the University of Pittsburgh. Pancreatic acini were harvested40–42 and preincubated with 50 μmol/L orlistat29 or cetilistat, after which linoleic acid (LA)29 or GTL was added, followed by incubation for 4 hours. At the end of which, cell death was measured by lactate dehydrogenase (LDH) leakage29 or glycerol generation was measured29 to quantify GTL hydrolysis.
Hypothermia slows sequential and parallel steps initiated during caerulein pancreatitis
2014, PancreatologyCitation Excerpt :This was then diluted and assayed in trypsin assay buffer with 25 mM calcium chloride at the same temperature. To determine cathepsin B mediated TGA, activity generated in the absence of cathepsin B at the same temperature was subtracted from the activity generated in the presence of cathepsin B. For trypsin measurements in acinar homogenates, these were prepared as described previously [2,3,35] using the buffer and substrate described above and depicted as arbitrary units per microgram of DNA in homogenate. The activities were measured spectrophotometrically (Bio-Versa, Bio-Rad).
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Address for reprint requests to: Michael L. Steer, M. D., Department of Surgery, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215. fax: (617) 667-8679; e-mail: [email protected].
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Supported by National Institutes of Health grant DK-31396.