Effect of cinnamtannin B‐1 on cholecystokinin‐8‐evoked responses in mouse pancreatic acinar cells

Abstract

1. Cinnamtannin B-1 is a naturally occurring A-type proanthocyanidin that belongs to a class of polyphenols widely distributed throughout the plant kingdom and exhibiting anti-oxidant properties. 2. In the present study, we examined the effects of cinnamtannin B-1 on cholecystokinin octapeptide (CCK-8)-evoked Ca(2+) mobilization, reactive oxygen species (ROS) production and amylase secretion in the exocrine pancreas. 3. Stimulation of cells with 1 nmol/L CCK-8 led to a transient increase in the cytosolic free calcium concentration ([Ca(2+) ](c) ), followed by a decrease towards a value close to the prestimulation level. In the presence of 10 μmol/L cinnamtannin B-1, stimulation of cells with CCK-8 resulted in a smaller [Ca(2+) ](c) peak response, a faster rate of decay of [Ca(2+) ](c) and lower values for the steady state of [Ca(2+) ](c) , compared with the effect of CCK-8 alone. Cinnamtannin B-1 decreased Ca(2+) influx after depletion of intracellular stores by either CCK-8 or thapsigargin (1 μmol/L). Conversely, CCK-8 increased the fluorescence of 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate acetyl ester (CM-H(2) DCFDA), reflecting an increase in oxidation. Cinnamtannin B-1 reduced CCK-8-induced oxidation of CM-H(2) DCFDA. Cholecystokinin-8 had a biphasic effect on amylase secretion, producing maximum at a concentration of 0.1 nmol/L and reducing secretion at higher concentrations. Pre-incubation of cells with 10 μmol/L cinnamtannin B-1 significantly attenuated the inhibition of enzyme secretion in response to high concentrations of CCK-8 (i.e. >10(-10) mol/L). Finally, the anti-oxidant protected acinar cells against CCK-8-induced cell death. 4. The beneficial effects of cinnamtannin B-1 appear to be mediated by a reduction in intracellular Ca(2+) overload, ROS production and intracellular accumulation of digestive enzymes, which is a common pathological precursor that mediates pancreatitis.