1. Volume 38, Issue 1, Pages 51-57 (January 2006)
Ethanol impairs CCK-8-evoked amylase secretion through Ca2+-mediated ROS generation in mouse pancreatic acinar cells 
Antonio González, Ana M. Núñez, María P. Granados, José A. Pariente, Ginés M. Salido 
Alcohol 
Volume 38, Issue 1, Pages (January 2006)
DOI: /j.alcohol
Copyright © 2006 Elsevier Inc. Terms and Conditions

2. Fig. 1
Effect of ethanol on amylase secretion. Pancreatic acinar cells were incubated with increasing concentrations of cholecystokinin octapeptide (CCK-8) (full squares), in the presence of 50mM ethanol (open circles) or in the presence of 50mM ethanol plus 2mM of the antioxidant dithiothreitol (open squares). Inset: pancreatic acinar cells were incubated with increasing concentrations of ethanol. Amylase activities were expressed as percent of total content of amylase at the beginning of the incubation as described in the Materials and methods section. Results are means±S.E.M. of 7–25 different experiments (pancreata). (†, P<.01 and ††, P<.001 with respect to CCK-8 alone. *, P<.05, **, P<.001, and ***, P<.001 with respect to CCK-8 plus ethanol.)
Alcohol  , 51-57DOI: ( /j.alcohol )
Copyright © 2006 Elsevier Inc. Terms and Conditions

3. Fig. 2
Time-course of fluorescence changes in fura-2-loaded pancreatic acinar cells. The graphics show the effect of ethanol on cytosolic free Ca2+ concentration ([Ca2+]c) in pancreatic acinar cells loaded with the fluorescent probe fura-2-AM. Changes in fluorescence emitted by this fluorophore reflect changes in [Ca2+]c. Stimulation of cells with 50mM ethanol led to a slow and sustained increase in [Ca2+]c over the basal (resting) level, both in the presence (A) and in the absence (B) of Ca2+ in the extracellular medium. The arrow indicates the moment of addition of ethanol to the cell suspension in the cuvette of a spectrofluorimeter. Traces represent the mean of four to five independent experiments (pancreata) (EtOH, ethanol).
Alcohol  , 51-57DOI: ( /j.alcohol )
Copyright © 2006 Elsevier Inc. Terms and Conditions

4. Fig. 3
Time-course of reactive oxygen species (ROS) production evoked by ethanol in mouse pancreatic acinar cells. (A) Stimulation of CM-H2DCFDA-loaded pancreatic acinar cells with 50mM ethanol led to an increase in ROS generation over the basal (resting) value (full circles). In the absence of extracellular Ca2+ and in the presence of 1mM of the Ca2+ chelator ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) (open circles) or in the additional presence of the cytosolic Ca2+ chelator BAPTA (10μM) (full squares), the generation of ROS upon addition of ethanol to the cells was decreased. The arrow indicates the moment of addition of ethanol to the cell suspension at a concentration to reach 50mM in the cuvette. Traces represent the mean of four to eight independent experiments (pancreata) (EtOH, ethanol). (B) Histogram depicting the values of fluorescence of CM-H2DCFDA-loaded pancreatic acinar cells in control conditions (full bar), following stimulation of cells with 50mM ethanol alone (vertical bars), in the absence of extracellular Ca2+ and in the presence of 1mM EGTA in the extracellular medium (horizontal bars), in the absence of extracellular Ca2+ plus 1mM EGTA and in the presence of the cytosolic Ca2+ chelator BAPTA (10μM) (open bar), and following addition of 1μM Tps plus 5μM ionomycin to the cell suspension (dotted bar). Graphics are representative of five to seven independent experiments (pancreata). Tps, thapsigargin; ion, ionomycin; *, P<.01 compared to basal; **, P<.001 compared to basal; †, P<.001 compared to ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) and EGTA plus BAPTA.
Alcohol  , 51-57DOI: ( /j.alcohol )
Copyright © 2006 Elsevier Inc. Terms and Conditions