Ethanol exerts dual effects on calcium homeostasis in CCK-8-stimulated mouse pancreatic acinar cells.
BACKGROUND
A significant percentage of patients with pancreatitis often presents a history of excessive alcohol consumption. Nevertheless, the patho-physiological effect of ethanol on pancreatitis remains poorly understood. In the present study, we have investigated the early effects of acute ethanol exposure on CCK-8-evoked Ca2+ signals in mouse pancreatic acinar cells. Changes in [Ca2+]i and ROS production were analyzed employing fluorescence techniques after loading cells with fura-2 or CM-H2DCFDA, respectively.


RESULTS
Ethanol, in the concentration range from 1 to 50 mM, evoked an oscillatory pattern in [Ca2+]i. In addition, ethanol evoked reactive oxygen species generation (ROS) production. Stimulation of cells with 1 nM or 20 pM CCK-8, respectively led to a transient change and oscillations in [Ca2+]i. In the presence of ethanol a transformation of 20 pM CCK-8-evoked physiological oscillations into a single transient increase in [Ca2+]i in the majority of cells was observed. Whereas, in response to 1 nM CCK-8, the total Ca2+ mobilization was significantly increased by ethanol pre-treatment. Preincubation of cells with 1 mM 4-MP, an inhibitor of alcohol dehydrogenase, or 10 microM of the antioxidant cinnamtannin B-1, reverted the effect of ethanol on total Ca2+ mobilization evoked by 1 nM CCK-8. Cinnamtannin B-1 blocked ethanol-evoked ROS production.


CONCLUSIONS
ethanol may lead, either directly or through ROS generation, to an over stimulation of pancreatic acinar cells in response to CCK-8, resulting in a higher Ca2+ mobilization compared to normal conditions. The actions of ethanol on CCK-8-stimulation of cells create a situation potentially leading to Ca2+ overload, which is a common pathological precursor that mediates pancreatitis.
Fernández-Sánchez
Marcela
M
Department of Physiology, Cell Physiology Research Group, University of Extremadura, Cáceres, Spain. marpacepelapa@hotmail.com
del Castillo-Vaquero
Angel
A
Salido
Ginés M
GM
González
Antonio
A
D000426:Alcohol Dehydrogenase / Q000378:metabolism; D000818:Animals; D000872:Anthocyanins / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D000431:Ethanol / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D016257:Fura-2 / Q000737:chemistry / Q000378:metabolism; D006706:Homeostasis / Q000187:drug effects; D008297:Male; D051379:Mice; D046790:Pancreas, Exocrine / Q000166:cytology / Q000378:metabolism*; D044945:Proanthocyanidins; D017382:Reactive Oxygen Species / Q000378:metabolism; D012844:Sincalide / Q000494:pharmacology*; D013997:Time Factors
D000872:Anthocyanins; D000975:Antioxidants; D005456:Fluorescent Dyes; D044945:Proanthocyanidins; D017382:Reactive Oxygen Species; C517872:cinnamtannin B-1; D000431:Ethanol; D000426:Alcohol Dehydrogenase; D012844:Sincalide; D002118:Calcium; D016257:Fura-2