Basic ResearchEthanol metabolism and transcription factor activation in pancreatic acinar cells in rats☆,**
Section snippets
Isolation of pancreatic acini
Pancreas was dissected from Sprague-Dawley rats (100-150 g), and dispersed pancreatic acini were isolated by a standard collagenase digestion method as described previously.9, 10, 12
Enzyme assays
Activities of the enzymes, FAEE synthase and ADH, were measured in cell or tissue lysates. To measure FAEE activity, tissue or isolated pancreatic acini were homogenized in 10 mmol/L Tris-HCl buffer (pH 8.0) containing 1 mmol/L phenylmethylsulfonyl fluoride, 1 mmol/L β-mercaptoethanol, and a protease-inhibitor
Characteristics of ethanol-metabolizing enzymes
The results in Figure 1 compare activities of ethanol-metabolizing enzymes in isolated pancreatic acini and pancreatic and liver tissue of normal rat.
Discussion
The results of this study show that pancreatic acinar cells are the main source of ethanol metabolism in the pancreas because isolated acinar cells and pancreatic tissue show the same levels of activities of ethanol-metabolizing enzymes. In the pancreas, accumulation of the oxidative metabolite, acetate, is greater than of the nonoxidative product, FAEE. The activity of ADH is also greater than that of FAEE synthase. Of note, the values for both the oxidative and nonoxidative pathways represent
Acknowledgements
The authors thank David T. Kira for help with nuclear extract preparations, and Yoon Jung for help in preparing this article.
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2020, Biochemical PharmacologyCitation Excerpt :FAEEs can be detected in plasma and other tissues after alcohol consumption [7,9]. On the other hand, pancreatic alcohol dehydrogenase (ADH) and cytochrome P450E1 (CYP2E1) activities involved in the canonical oxidative pathway of EtOH metabolism are relatively low or negligible [5]. Inhibition of hepatic ADH1 (a major enzyme involved in EtOH oxidation) leads to increased biosynthesis of FAEEs in the pancreas and toxicity to the pancreatic acinar cells [2,5,6,8,10–15].
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Address requests for reprints to: Anna S. Gukovskaya, Ph.D., Veterans Affairs Greater Los Angeles Healthcare System, Building 258, Room 340, 11301 Wilshire Boulevard, Los Angeles, California 90073. e-mail: [email protected]; fax: (310) 268-4578.
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Supported by the Research Center for Alcoholic Liver and Pancreatic Diseases grant (P50-A11999) from the National Institute on Alcohol Abuse and Alcoholism. The gas chromatography–mass spectrometry measurements were supported by a Veterans Administration Merit Review Award (to L.D.F.).