CONCLUSION: Since phosphatidic acid (PA), a product of phospholipase D(PLD), is known as a second messenger probably involved in cell proliferation and differentiation, our results potentially suggest a new mechanism for pancreatic tissue injury after ethanol ingestion. BACKGROUND: The mechanisms by which ethanol causes pancreatic injury are still not clear. In vitro studies have suggested a relationship of PLD to ethanol metabolism. This study was undertaken to establish the involvement of PLD in ethanol metabolism in isolated pancreatic acini and to determine the influence of acute ethanol ingestion on PLD activity in pancreas and pancreatic growth after cerulein (Ce) infusion. METHODS: Dispersed pancreatic acini prelabeled with 3H myristic acid were incubated with 500 pM Ce in the presence of different concentrations of ethanol; then labeled PA and phosphatidylethanol (PEt) production were measured under the same experimental conditions. For in vivo study, male rats were infused with Ce (0.25 microgram/kg/h) or saline; 1 h before infusion, animals were treated with 40% ethanol (5 g/kg p.o.) or saline, respectively. After 1, 2, and 48 h of Ce infusion, rats were killed; dispersed pancreatic acini were then prepared and PLD activity was measured. Pancreatic weight, protein, RNA, and DNA content were also established. RESULTS: The production of PEt in vitro after Ce stimulation was significantly elevated with 1% ethanol in the medium. In the presence of different concentrations of ethanol (0.5-2%), a significant inhibition of PA accumulation in in vitro experiments was observed. The decrease of PA accumulation with ethanol was parallel to the increase of PEt production under the same experimental conditions. PLD activity was significantly elevated after 1 and 2 h of Ce infusion (116 and 105%, respectively), reaching control value after 48 h. Acute ethanol ingestion significantly diminished PLD activity after 1 and 2 h. After 48 h of Ce infusion, a significant increase in pancreatic weight, protein, RNA, and DNA content in pancreatic tissue was found. Ethanol was not able to influence pancreatic weight, proteins and RNA content. However, it had the potency to diminish DNA content after 48 h of Ce infusion.
CONCLUSION: Since phosphatidic acid (PA), a product of phospholipase D(PLD), is known as a second messenger probably involved in cell proliferation and differentiation, our results potentially suggest a new mechanism for pancreatic tissue injury after ethanol ingestion. BACKGROUND: The mechanisms by which ethanol causes pancreatic injury are still not clear. In vitro studies have suggested a relationship of PLD to ethanol metabolism. This study was undertaken to establish the involvement of PLD in ethanol metabolism in isolated pancreatic acini and to determine the influence of acute ethanol ingestion on PLD activity in pancreas and pancreatic growth after cerulein (Ce) infusion. METHODS: Dispersed pancreatic acini prelabeled with 3H myristic acid were incubated with 500 pM Ce in the presence of different concentrations of ethanol; then labeled PA and phosphatidylethanol (PEt) production were measured under the same experimental conditions. For in vivo study, male rats were infused with Ce (0.25 microgram/kg/h) or saline; 1 h before infusion, animals were treated with 40% ethanol (5 g/kg p.o.) or saline, respectively. After 1, 2, and 48 h of Ce infusion, rats were killed; dispersed pancreatic acini were then prepared and PLD activity was measured. Pancreatic weight, protein, RNA, and DNA content were also established. RESULTS: The production of PEt in vitro after Ce stimulation was significantly elevated with 1% ethanol in the medium. In the presence of different concentrations of ethanol (0.5-2%), a significant inhibition of PA accumulation in in vitro experiments was observed. The decrease of PA accumulation with ethanol was parallel to the increase of PEt production under the same experimental conditions. PLD activity was significantly elevated after 1 and 2 h of Ce infusion (116 and 105%, respectively), reaching control value after 48 h. Acute ethanol ingestion significantly diminished PLD activity after 1 and 2 h. After 48 h of Ce infusion, a significant increase in pancreatic weight, protein, RNA, and DNA content in pancreatic tissue was found. Ethanol was not able to influence pancreatic weight, proteins and RNA content. However, it had the potency to diminish DNA content after 48 h of Ce infusion.