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Ethanol and fatty acids strongly decrease the activity and the expression of CFTR CL− channel in pancreatic ductal epithelial cell
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Abstracts / Pancreatology 13 (2013) S2–S98
Introduction: Fluid and HCO 3 secretion is a vital function of pancreatic ductal eptihelia and their role in acute pancreatitis (AP) is poorly characterized. NHERF-1 is a cytosolic scaffolding protein mediating the apical targeting and retention of ion channels and transporters, such as cystic fibrosis transmembrane conductance regulator (CFTR). Aims: The main aims of this study were to investigate the physiological and pathophysiological relevance of NHERF-1 expression in the pancreas. Materials & methods: We analyzed the effects of NHERF-1 deletion on ductal function both in vitro and in vivo. The localization of CFTR in wildtype and NHERF-1 KO mice was performed by immunohistochemistry. AP was induced by administration of intraperitoneal cerulein or by intraductal sodium-taurocholate. The severity of AP was evaluated by measureing histological and laboratory parameters. Results: NHERF-1 mRNA was markedly expressed in the pancreatic ducts of wild-type mice. We show that NHERF-1 plays a critical role in modulating the apical localization of pancreatic ductal CFTR. The translocalization of CFTR resulted in significantly lower pancreatic ductal bicarbonate and fluid secretion. NHERF-1 expression also influenced the development of AP in both mouse models; the disease severity, especially the degree of acinar cell death, was higher in NHERF-1-knock-out vs. wildtype mice. Conclusion: Our findings provide evidence for the crucial role of ductal fluid and HCO-3 secretion in the protection of pancreas from acute stressors, which cause AP. € AMOP. This study was supported by OTKA, MTA/DFG and NFU/T
O-11 Abstract id: 281. The crucial role of ATPi in the inhibitory effect of ethanol and its nonoxidative metabolites on CFTR in pancreatic ductal cells k 2, Zolta n Rakonczay, Jr. 2, Jo zsef Viktoria Venglovecz 1, Linda Juda eter Hegyi 2. Mal eth 2, A. Mike Gray 3, P 1 Department of Pharmacology and Pharmacotherapy, University of Szeged, Hungary 2 First Department of Medicine, University of Szeged, Hungary 3 Institute for Cell and Molecular Biosciences, University Medical School, Newcastle upon Tyne, United Kingdom
Introduction: Excessive alcohol consumption causes acute pancreatitis but the mechanism involved is not well understood. Recent investigations suggest that pancreatic ductal epithelial cells (PDECs) are involved in the pathogenesis of pancreatitis. The cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel plays a major role in PDEC anion and fluid secretion, and that dysfunction of CFTR is often associated with pancreatitis. Aims: Our aim was to investigate the effect of ethanol and its nonoxidative metabolites on CFTR activity. Materials & methods: The dose- and time-dependent effects of ethanol, its oxidative and nonoxidative metabolites (acetaldehyde (Ac) and palmitoleic acid ethyl ester (POAEE), respectively) and palmitoleic acid (POA) were investigated on CFTR activity on freshly isolated guinea pig PDECs and Capan-1 cell line, using the whole cell configuration of the patch clamp technique. Changes in intracellular ATP (ATPi) were measured by spectrofluorometry. Results: Ethanol (10 and 100 mM) significantly increased the basal, but reversibly blocked forskolin-stimulated CFTR currents. The inhibitory effect of ethanol was mimicked by POAEE and POA, the latter being produced by fatty acid ethyl esterase hydrolase. Ethanol, POAEE and POA caused depletion of ATPi linked to CFTR inhibition, since their inhibitory effects were almost completely abolished if ATPi depletion was prevented. Conclusion: We propose that ethanol causes functional damage of CFTR through an ATPi-dependent mechanism. Furthermore, we suggest that the maintenance of ductal ATPi may represent a therapeutic option in the treatment of the disease. € AMOP. This study was supported by OTKA, MTA and NFU/T
O-12 Abstract id: 273. Ethanol and fatty acids strongly decrease the activity and the expression of CFTR CLL channel in pancreatic ductal epithelial cell zs 1, Petra Pallagi 1, Linda Juda k 2, Lajos zsef Mal Jo eth 1, Anita Bala cz 3, Katalin Borka 3, Vikto n ria Venglovecz 2, 3, Zolta Somora Kem eny 1, Aron eter Hegyi 1. Rakonczay 1, Mike Gray 4, P 1
First Dept. of Medicine, University of Szeged, Szeged, Hungary Dept. of Pharmacology and Pharmacotherapy, University of Szeged, Szeged, Hungary 3 II. Dept. of Pathology, Semmelweis University, Budapest, Hungary 4 Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle, United Kingdom 2
Introduction: Excessive ethanol consumption is one of the most common causes of acute pancreatitis. Sarles observed elevated sweat Cl- concentration in alcoholic patients, which suggests decreased function of cystic fibrosis transmembrane conductance regulator (CFTR), however the effect of ethanol on pancreatic bicarbonate secretion was not investigated in details. Aims: Our aim was to evaluate the effects of ethanol and non-oxidative ethanol metabolites on pancreatic ductal epithelial cells (PDEC). Materials & methods: In our experiments Capan-1 cells, guinea pig PDEC and human pancreatic tissue were used. The effects of ethanol, fatty acid ethyl esters and fatty acids on intracellular pH (pHi), Ca2þ concentration ([Ca2þ]i), ATP [(ATP)i] and CFTR Cl- current of PDEC were measured. The expression and localization of CFTR were detected in PDEC and in human pancreatic tissue. Results: The administration of 10mM ethanol stimulated pancreatic HCO-3 secretion via IP3 mediated [Ca2þ]i elevation. In contrast, 100mM EtOH and 200mM palmitoleic acid (POA) inhibited the HCO-3 secretion of PDEC and decreased the CFTR Cl- current via sustained [Ca2þ]i elevation and (ATP)i depletion. We also showed that ethanol and POA significantly decreased the expression of CFTR in PDEC after 48h incubation. Moreover, the CFTR expression of intralobular pancreatic ducts was significantly decreased in acute (AP) and chronic pancreatitis (CP) patients. Conclusion: These results suggest that CFTR could play an important role in the pathogenesis of alcohol induced pancreatitis. Restoration of CFTR localization and function may be potential therapeutic possibility in alcohol induced AP and CP. € AMOP. This work was supported by OTKA, MTA and NFU/T
O-13 Best of APA. Lessons on aggressive intravenous hydration in acute pancreatitis: A meta-analysis of clinical trials Rabin Rahmani MD, Daniel Marino MS, Steven Shamah MD, Ira Mayer MD, Scott Tenner MD, MPH. Division of Gastroenterology, Department of Medicine, State University of New York, Health Sciences Center, Brooklyn, NY, USA Despite limited clinical research in humans, over the past decade, a dozen guidelines from the United States and Europe on the management of acute pancreatitis have put forth aggressive intravenous hydration at the forefront. Largely based on animal studies and indirect evidence in humans, experts have put forth the notion that aggressive intravenous hydration, early in the course, would attenuate the severity of the disease in patients with acute pancreatitis. Recent clinical studies in human subjects evaluating the efficacy of aggressive intravenous hydration have demonstrated conflicting results. The purpose of our study was to systematically review, analyze and combine the different studies in a metaanalysis to determine patterns of efficacy that may exist in order to promote appropriate fluid management in patients with acute pancreatitis. In order to be included in the analysis, the published study needed to include patients with acute pancreatitis were enrolled in a consecutive, prospective or retrospective fashion. The study needed to provide information regarding the amount of fluid for each group, and outcome,
Abstracts / Pancreatology 13 (2013) S2–S98
Introduction: Fluid and HCO 3 secretion is a vital function of pancreatic ductal eptihelia and their role in acute pancreatitis (AP) is poorly characterized. NHERF-1 is a cytosolic scaffolding protein mediating the apical targeting and retention of ion channels and transporters, such as cystic fibrosis transmembrane conductance regulator (CFTR). Aims: The main aims of this study were to investigate the physiological and pathophysiological relevance of NHERF-1 expression in the pancreas. Materials & methods: We analyzed the effects of NHERF-1 deletion on ductal function both in vitro and in vivo. The localization of CFTR in wildtype and NHERF-1 KO mice was performed by immunohistochemistry. AP was induced by administration of intraperitoneal cerulein or by intraductal sodium-taurocholate. The severity of AP was evaluated by measureing histological and laboratory parameters. Results: NHERF-1 mRNA was markedly expressed in the pancreatic ducts of wild-type mice. We show that NHERF-1 plays a critical role in modulating the apical localization of pancreatic ductal CFTR. The translocalization of CFTR resulted in significantly lower pancreatic ductal bicarbonate and fluid secretion. NHERF-1 expression also influenced the development of AP in both mouse models; the disease severity, especially the degree of acinar cell death, was higher in NHERF-1-knock-out vs. wildtype mice. Conclusion: Our findings provide evidence for the crucial role of ductal fluid and HCO-3 secretion in the protection of pancreas from acute stressors, which cause AP. € AMOP. This study was supported by OTKA, MTA/DFG and NFU/T
O-11 Abstract id: 281. The crucial role of ATPi in the inhibitory effect of ethanol and its nonoxidative metabolites on CFTR in pancreatic ductal cells k 2, Zolta n Rakonczay, Jr. 2, Jo zsef Viktoria Venglovecz 1, Linda Juda eter Hegyi 2. Mal eth 2, A. Mike Gray 3, P 1 Department of Pharmacology and Pharmacotherapy, University of Szeged, Hungary 2 First Department of Medicine, University of Szeged, Hungary 3 Institute for Cell and Molecular Biosciences, University Medical School, Newcastle upon Tyne, United Kingdom
Introduction: Excessive alcohol consumption causes acute pancreatitis but the mechanism involved is not well understood. Recent investigations suggest that pancreatic ductal epithelial cells (PDECs) are involved in the pathogenesis of pancreatitis. The cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel plays a major role in PDEC anion and fluid secretion, and that dysfunction of CFTR is often associated with pancreatitis. Aims: Our aim was to investigate the effect of ethanol and its nonoxidative metabolites on CFTR activity. Materials & methods: The dose- and time-dependent effects of ethanol, its oxidative and nonoxidative metabolites (acetaldehyde (Ac) and palmitoleic acid ethyl ester (POAEE), respectively) and palmitoleic acid (POA) were investigated on CFTR activity on freshly isolated guinea pig PDECs and Capan-1 cell line, using the whole cell configuration of the patch clamp technique. Changes in intracellular ATP (ATPi) were measured by spectrofluorometry. Results: Ethanol (10 and 100 mM) significantly increased the basal, but reversibly blocked forskolin-stimulated CFTR currents. The inhibitory effect of ethanol was mimicked by POAEE and POA, the latter being produced by fatty acid ethyl esterase hydrolase. Ethanol, POAEE and POA caused depletion of ATPi linked to CFTR inhibition, since their inhibitory effects were almost completely abolished if ATPi depletion was prevented. Conclusion: We propose that ethanol causes functional damage of CFTR through an ATPi-dependent mechanism. Furthermore, we suggest that the maintenance of ductal ATPi may represent a therapeutic option in the treatment of the disease. € AMOP. This study was supported by OTKA, MTA and NFU/T
O-12 Abstract id: 273. Ethanol and fatty acids strongly decrease the activity and the expression of CFTR CLL channel in pancreatic ductal epithelial cell zs 1, Petra Pallagi 1, Linda Juda k 2, Lajos zsef Mal Jo eth 1, Anita Bala cz 3, Katalin Borka 3, Vikto n ria Venglovecz 2, 3, Zolta Somora Kem eny 1, Aron eter Hegyi 1. Rakonczay 1, Mike Gray 4, P 1
First Dept. of Medicine, University of Szeged, Szeged, Hungary Dept. of Pharmacology and Pharmacotherapy, University of Szeged, Szeged, Hungary 3 II. Dept. of Pathology, Semmelweis University, Budapest, Hungary 4 Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle, United Kingdom 2
Introduction: Excessive ethanol consumption is one of the most common causes of acute pancreatitis. Sarles observed elevated sweat Cl- concentration in alcoholic patients, which suggests decreased function of cystic fibrosis transmembrane conductance regulator (CFTR), however the effect of ethanol on pancreatic bicarbonate secretion was not investigated in details. Aims: Our aim was to evaluate the effects of ethanol and non-oxidative ethanol metabolites on pancreatic ductal epithelial cells (PDEC). Materials & methods: In our experiments Capan-1 cells, guinea pig PDEC and human pancreatic tissue were used. The effects of ethanol, fatty acid ethyl esters and fatty acids on intracellular pH (pHi), Ca2þ concentration ([Ca2þ]i), ATP [(ATP)i] and CFTR Cl- current of PDEC were measured. The expression and localization of CFTR were detected in PDEC and in human pancreatic tissue. Results: The administration of 10mM ethanol stimulated pancreatic HCO-3 secretion via IP3 mediated [Ca2þ]i elevation. In contrast, 100mM EtOH and 200mM palmitoleic acid (POA) inhibited the HCO-3 secretion of PDEC and decreased the CFTR Cl- current via sustained [Ca2þ]i elevation and (ATP)i depletion. We also showed that ethanol and POA significantly decreased the expression of CFTR in PDEC after 48h incubation. Moreover, the CFTR expression of intralobular pancreatic ducts was significantly decreased in acute (AP) and chronic pancreatitis (CP) patients. Conclusion: These results suggest that CFTR could play an important role in the pathogenesis of alcohol induced pancreatitis. Restoration of CFTR localization and function may be potential therapeutic possibility in alcohol induced AP and CP. € AMOP. This work was supported by OTKA, MTA and NFU/T
O-13 Best of APA. Lessons on aggressive intravenous hydration in acute pancreatitis: A meta-analysis of clinical trials Rabin Rahmani MD, Daniel Marino MS, Steven Shamah MD, Ira Mayer MD, Scott Tenner MD, MPH. Division of Gastroenterology, Department of Medicine, State University of New York, Health Sciences Center, Brooklyn, NY, USA Despite limited clinical research in humans, over the past decade, a dozen guidelines from the United States and Europe on the management of acute pancreatitis have put forth aggressive intravenous hydration at the forefront. Largely based on animal studies and indirect evidence in humans, experts have put forth the notion that aggressive intravenous hydration, early in the course, would attenuate the severity of the disease in patients with acute pancreatitis. Recent clinical studies in human subjects evaluating the efficacy of aggressive intravenous hydration have demonstrated conflicting results. The purpose of our study was to systematically review, analyze and combine the different studies in a metaanalysis to determine patterns of efficacy that may exist in order to promote appropriate fluid management in patients with acute pancreatitis. In order to be included in the analysis, the published study needed to include patients with acute pancreatitis were enrolled in a consecutive, prospective or retrospective fashion. The study needed to provide information regarding the amount of fluid for each group, and outcome,