Darbaz Awla1, Anna V Zetterqvist2, Aree Abdulla1, Cristina Camello3, Lisa M Berglund2, Peter Spégel4, Maria J Pozo3, Pedro J Camello3, Sara Regnér1, Maria F Gomez2, Henrik Thorlacius5. 1. Department of Clinical Sciences, Section of Surgery, Skåne University Hospital, Malmö, Sweden. 2. Department of Clinical Sciences, Vascular Excitation-Transcription Coupling, Lund University, Malmö, Sweden. 3. Department of Physiology, Nursing School, University of Extremadura, Caceres, Spain. 4. Department of Clinical Sciences, Molecular Metabolism, Lund University, Malmö, Sweden. 5. Department of Clinical Sciences, Section of Surgery, Skåne University Hospital, Malmö, Sweden. Electronic address: henrik.thorlacius@med.lu.se.
Abstract
BACKGROUND & AIMS: The signaling mechanisms that regulate trypsinogen activation and inflammation in acute pancreatitis (AP) are unclear. We explored the involvement of the calcium- and calcineurin-dependent transcription factor nuclear factor of activated T cells (NFAT) in development of AP in mice. METHODS: We measured levels of myeloperoxidase and macrophage inflammatory protein 2 (CXCL2), trypsinogen activation, and tissue damage in the pancreas 24 hours after induction of AP by retrograde infusion of taurocholate into the pancreatic ducts of wild-type, NFAT luciferase reporter (NFAT-luc), and NFATc3-deficient mice. We isolated acinar cells and measured NFAT nuclear accumulation, trypsin activity, and expression of NFAT-regulated genes. RESULTS: Infusion of taurocholate increased the transcriptional activity of NFAT in the pancreas, aorta, lung, and spleen of NFAT-luc mice. Inhibition of NFAT with A-285222 blocked taurocholate-induced activation of NFAT in all organs. A-285222 also reduced taurocholate-induced increases in levels of amylase, myeloperoxidase, and CXCL2; activation of trypsinogen; necrosis of acinar cells; edema; leukocyte infiltration; and hemorrhage in the pancreas. NFATc3-deficient mice were protected from these effects of taurocholate. Similar results were obtained using an l-arginine-induced model of AP. Reverse-transcription polymerase chain reaction and confocal immunofluorescence analyses showed that NFATc3 is expressed by acinar cells. NFATc3 expression was activated by stimuli that increase intracellular calcium levels, and activation was prevented by the calcineurin blocker cyclosporin A or A-285222. Activation of trypsinogen by secretagogues in acinar cells was prevented by pharmacologic inhibition of NFAT signaling or lack of NFATc3. A-285222 also reduced expression of inflammatory cytokines such as CXCL2 in acinar cells. CONCLUSIONS: NFATc3 regulates trypsinogen activation, inflammation, and pancreatic tissue damage during development of AP in mice and might be a therapeutic target.
BACKGROUND & AIMS: The signaling mechanisms that regulate trypsinogen activation and inflammation in acute pancreatitis (AP) are unclear. We explored the involvement of the calcium- and calcineurin-dependent transcription factor nuclear factor of activated T cells (NFAT) in development of AP in mice. METHODS: We measured levels of myeloperoxidase and macrophage inflammatory protein 2 (CXCL2), trypsinogen activation, and tissue damage in the pancreas 24 hours after induction of AP by retrograde infusion of taurocholate into the pancreatic ducts of wild-type, NFAT luciferase reporter (NFAT-luc), and NFATc3-deficient mice. We isolated acinar cells and measured NFAT nuclear accumulation, trypsin activity, and expression of NFAT-regulated genes. RESULTS: Infusion of taurocholate increased the transcriptional activity of NFAT in the pancreas, aorta, lung, and spleen of NFAT-luc mice. Inhibition of NFAT with A-285222 blocked taurocholate-induced activation of NFAT in all organs. A-285222 also reduced taurocholate-induced increases in levels of amylase, myeloperoxidase, and CXCL2; activation of trypsinogen; necrosis of acinar cells; edema; leukocyte infiltration; and hemorrhage in the pancreas. NFATc3-deficient mice were protected from these effects of taurocholate. Similar results were obtained using an l-arginine-induced model of AP. Reverse-transcription polymerase chain reaction and confocal immunofluorescence analyses showed that NFATc3 is expressed by acinar cells. NFATc3 expression was activated by stimuli that increase intracellular calcium levels, and activation was prevented by the calcineurin blocker cyclosporin A or A-285222. Activation of trypsinogen by secretagogues in acinar cells was prevented by pharmacologic inhibition of NFAT signaling or lack of NFATc3. A-285222 also reduced expression of inflammatory cytokines such as CXCL2 in acinar cells. CONCLUSIONS:NFATc3 regulates trypsinogen activation, inflammation, and pancreatic tissue damage during development of AP in mice and might be a therapeutic target.
Authors: Yaser Panahi; Shohreh Fakhari; Mehdi Mohammadi; Mohammad Reza Rahmani; Mohammad Saeid Hakhamaneshi; Ali Jalili Journal: Am J Clin Exp Immunol Date: 2015-07-05
Authors: Kamaldeen A Muili; Dong Wang; Abrahim I Orabi; Sheharyar Sarwar; Yuhuan Luo; Tanveer A Javed; John F Eisses; Syeda M Mahmood; Shunqian Jin; Vijay P Singh; Meena Ananthanaravanan; George Perides; John A Williams; Jeffery D Molkentin; Sohail Z Husain Journal: J Biol Chem Date: 2012-11-12 Impact factor: 5.157
Authors: Balázs Kui; Zsolt Balla; Eszter T Végh; Petra Pallagi; Viktória Venglovecz; Béla Iványi; Tamás Takács; Péter Hegyi; Zoltán Rakonczay Journal: Lab Invest Date: 2013-12-23 Impact factor: 5.662