Substance p regulates chemokine production in leukocytes implications in the pathogenesis of acute pancreatitis and associated lung injury

ACKNOWLEDGEMENTS First of all, I would like to express my heartfelt gratitude to my supervisor, Associate Professor Madhav Bhatia for providing me the great opportunity to work on this project and for his invaluable advice, guidance, continuous support and encouragement throughout the study I would also like to thank Ms Shoon Mei Leng, the senior laboratory officer, for her assistance, support and her excellent work in maintaining our orderly conducive laboratory environment My appreciation is also extended to every member of the Life Sciences Institute Cardiovascular Biology Program, Akhil Hegde, Cao Yang, He Min, Jenab Nooruddinbhai Sidhapuriw, Koh Yung Hua, Raina Devi Ramnath, Ramasamy Tamizhselvi, Selena Sio Weishan, and Zhi Liang for insightful discussions, technical advice and help in one way or another Last but not least, I would like to give my special thanks to my parents Without their support and encouragement, this project could not have been completed i    TABLE OF CONTENTS ACKNOWLEDGEMENTS I SUMMARY VI LIST OF FIGURES IX ABBREVIATIONS XIII PUBLICATIONS XV CHAPTER I INTRODUCTION 1.1 GENERAL OVERVIEW .1 1.2 ACUTE PANCREATITIS 1.2.1 Pathophysiology of acute pancreatitis and associated lung injury 1.2.1.1 Inflammatory mediators in acute pancreatitis and associated lung injury 1.2.1.2 Leukocytes in acute pancreatitis and associated lung injury 1.2.1.2.1 Leukocyte trafficking during inflammation 1.2.1.2.2 Neutrophils 1.2.1.2.3 Macrophages 1.2.2 Experimental models of acute pancreatitis 1.2.2.1 Principle of caerulein-induced acute pancreatitis model 1.2.2.2 Induction and characteristics of caerulein-induced acute pancreatitis model .10 1.3 SP 11 1.3.1 SP in acute pancreatitis and associated lung injury 11 1.3.2 SP in immunoregulation 12 1.3.2.1 SP in immunoregulation: neutrophils 12 1.3.2.2 SP in immunoregulation: macrophages 13 1.4 CHEMOKINES .14 1.4.1 Chemokines and their receptors in acute pancreatitis and associated lung injury .15 1.5 INTRACELLULAR SIGNALING MOLECULES 16 1.5.1 NF-κB 17 1.5.2 MAPKs .18 1.5.3 PKC isoforms .19 1.5.4 PI3K-Akt 19 1.6 RESEARCH RATIONALE AND OBJECTIVES .21 1.6.1 Research rationale .21 1.6.2 Objectives 22 CHAPTER II: SP REGULATES CHEMOKINE EXPRESSION IN LEUKOCYTES DURING ACUTE PANCREATITIS AND ASSOCIATED LUNG INJURY 24 2.1 INTRODUCTION 24 2.2 MATERIALS AND METHODS 25 2.2.1 Animals 25 2.2.2 Reagents 26 2.2.3 Caerulein-induced acute pancreatitis and associated lung injury model 26 2.2.4 Total RNA isolation and RT-PCR 27 ii    2.2.5 Immunohistochemistry .28 2.2.6 Statistical analysis .29 2.3 RESULTS 30 2.3.1 Effects of caerulein hyperstimulation and CP96,345 treatment on pancreatic and lung MCP-1 expression .30 2.3.2 Effects of caerulein hyperstimulation and CP96,345 treatment on pancreatic and lung RANTES expression .30 2.3.3 Effects of caerulein hyperstimulation and CP96,345 treatment on pancreatic and lung MIP-1α expression .31 2.3.4 Effects of caerulein hyperstimulation and CP96,345 treatment on pancreatic and lung MIP-2 expression .31 2.3.5 Effects of caerulein hyperstimulation and CP96,345 treatment on immunohistochemical localization of MCP-1, MIP-1α and MIP-2 in the pancreas and lungs 32 2.4 DISCUSSION 32 CHAPTER III IN VITRO EVIDENCE OF INDUCTION OF CHEMOKINE PRODUCTION AND CHEMOKINE RECEPTOR EXPRESSION IN MOUSE NEUTROPHILS BY SP AND THE CELLULAR MECHANISM 46 3.1 INTRODUCTION 46 3.2 MATERIALS AND METHODS 47 3.2.1 Animals 47 3.2.2 Reagents 47 3.2.3 Isolation of mouse primary neutrophils .48 3.2.4 Cell treatment 49 3.2.5 Cell migration assay 49 3.2.6 Total RNA isolation and RT-PCR 50 3.2.7 ELISA 51 3.2.8 Flow cytometry 51 3.2.9 Immunofluorescence staining 51 3.2.10 Nuclear extract preparation 52 3.2.11 NF-κB DNA-binding activity assay 52 3.2.12 Whole cell lysate preparation and Western blot analysis 53 3.2.13 Statistical analysis .54 3.3 RESULTS 54 3.3.1 SP treatment activated mouse primary neutrophils 54 3.3.2 SP treatment activated the transcription factor NF-κB in mouse primary neutrophils .55 3.3.3 SP treatment enhanced chemokine expression in mouse primary neutrophils 56 3.3.4 SP treatment induced chemokine receptor expression in mouse primary neutrophils 57 3.3.5 SP treatment enhanced chemotactic responses of mouse primary neutrophils to rMIP-2 and rMIP-1α 58 3.4 DISCUSSION 59 CHAPTER IV IN VITRO EVIDENCE OF INDUCTION OF CHEMOKINE PRODUCTION IN MOUSE MACROPHAGES BY SP AND THE CELLULAR MECHANISMS – ROLE OF NF-KB AND ERK1/2 .72 4.1 INTRODUCTION 72 4.2 MATERIALS AND METHODS 73 4.2.1 Reagents 73 iii    4.2.2 Cell culture and treatment 73 4.2.3 Isolation of peritoneal macrophages 74 4.2.4 ELISA 74 4.2.5 Whole cell lysate preparation and Western blot analysis 75 4.2.6 Nuclear extract preparation 75 4.2.7 NF-κB DNA-binding activity assay .75 4.2.8 EMSA 75 4.2.9 Statistical analysis .76 4.3 RESULTS 76 4.3.1 SP induced chemokine production in mouse macrophages 76 4.3.2 NK-1R but not NK-2R antagonists abolished SP-induced chemokine production in mouse macrophages .78 4.3.3 SP enhanced NF-κB but not AP-1 activity via the NK-1R in mouse macrophages .78 4.3.4 SP activated NF-κB via the classical and atypical pathways in mouse macrophages .79 4.3.5 Inhibition of NF-κB activation abolished SP-induced chemokine production in mouse macrophages .80 4.3.6 SP activated ERK1/2 in mouse macrophages 80 4.3.7 Inhibition of ERK1/2 activation attenuated SP-induced chemokine production in mouse macrophages .81 4.3.8 Inhibition of ERK1/2 activation prevented SP-induced phosphorylation and nuclear translocation of NF-κB p65 in mouse macrophages 81 4.4 DISCUSSION 82 CHAPTER V THE CELLULAR MECHANISMS OF SP-INDUCED CHEMOKINE PRODUCTION IN MOUSE MACROPHAGES – ROLE OF PKC ISOFORMS AND PI3K-AKT 101 5.1 INTRODUCTION 101 5.2 MATERIALS AND METHODS 101 5.2.1 Reagents 101 5.2.2 Cell culture and treatment 102 5.2.3 Isolation of peritoneal macrophages 102 5.2.4 ELISA .102 5.2.5 Whole cell lysate preparation and Western blot analysis 102 5.2.6 Subcellular fractionation 102 5.2.7 Nuclear extract preparation 103 5.2.8 NF-κB DNA-binding activity assay 103 5.2.9 Statistical analysis .103 5.3 RESULTS 103 5.3.1 SP induced PKCα, δ and ε activation via NK-1R in mouse macrophages .103 5.3.2 PKCα, δ and ε were involved in SP-induced chemokine production in mouse macrophages 104 5.3.3 PKCα, δ and ε were involved in SP-induced ERK1/2 activation in mouse macrophages 105 5.3.4 PKCα, δ and ε were involved in SP-induced NF-κB activation in mouse macrophages 106 5.3.5 SP induced activation of PI3K-Akt pathway via NK-1R in mouse macrophages 106 5.3.6 PI3K-Akt pathway was involved in SP-induced ERK1/2 and NF-κB activation and chemokine production in mouse macrophages 107 5.3.7 PKCs and PI3K-Akt were two independent convergent pathways activated by SP in mouse macrophages .108 5.3.8 SP induced PKC and PI3K-Akt pathways in primary peritoneal macrophages 108 5.3.9 Signal transduction pathways induced by SP in mouse macrophages .109 iv    5.4 DISCUSSION .109 CHAPTER VI NKA INDUCES CHEMOKINE PRODUCTION BY MOUSE MACROPHAGES VIA ERK1/2- AND PI3K-AKT-NFKB PATHWAYS 139 6.1 INTRODUCTION 139 6.2 MATERIALS AND METHODS 140 6.2.1 Reagents 140 6.2.2 Cell treatment 141 6.2.3 Isolation of primary peritoneal macrophages 141 6.2.4 Whole cell lysate preparation and Western blot analysis 141 6.2.5 Nuclear extract preparation 141 6.2.6 NF-κB DNA-binding activity assay 142 6.2.7 Total RNA isolation and RT-PCR 142 6.2.8 Immunofluorescence staining 142 6.2.9 ELISA .143 6.2.10 Statistical analysis 143 6.3 RESULTS 143 6.3.1 NKA upregulated NK-1R expression in mouse macrophages lacking detectable NK-2R 143 6.3.2 NKA induced chemokine production via NK-1R in mouse macrophages 144 6.3.3 NKA induced NF-κB activation via NK-1R in mouse macrophages 145 6.3.4 NKA activated ERK1/2 and PI3K-Akt pathways in mouse macrophages 146 6.3.5 ERK1/2 and PI3K-Akt pathways were involved in NKA-induced NF-κB activation and chemokine production in mouse macrophages 146 6.4 DISCUSSION .147 CHAPTER VII SUMMARY OF CONTRIBUTIONS AND FUTURE RESEARCH 171 7.1 SUMMARY OF CONTRIBUTIONS 171 7.2 FUTURE RESEARCH 174 REFERENCES 177 v    SUMMARY The neuropeptide substance P (SP) is a well-recognized inflammatory mediator in acute pancreatitis However, the mechanism remains elusive Aiming to delineate its mechanistic actions, the present study attempts to trace the signal transduction cascade activated by SP in the regulation of release of inflammatory molecules that contribute to disease progression In a mouse model of caerulein-induced acute pancreatitis and associated lung injury, blockade of the biological actions of SP with a specific antagonist of its neurokinin-1 receptor (NK-1R) significantly attenuated the chemokine (MCP-1, MIP-1α, and MIP-2) expression in pancreas and lungs, which correlated with less leukocyte infiltration and alleviated tissue damages Resident/infiltrating leukocytes (neutrophils and macrophages) were identified as major chemokine-expressing cells in both pancreas and lungs These preliminary observations suggested a potential mechanistic connection between SP, leukocytes and chemokines in acute pancreatitis – SP acting via resident/recruited leukocytes in tissues to induce the release of chemokines, which further aggravate the condition In vitro studies on isolated cultured mouse neutrophils and macrophages supported that SP, via NK-1R, exerts a direct stimulatory effect on chemokine (MCP-1, MIP-1α, and MIP-2) production by both cell types In neutrophils, SP-NK-1R also induced the expression of CD11b (a neutrophil activation marker) and chemokine receptors (CCR1 and CXCR2) As a functional consequence, SP-stimulated neutrophils exhibited vi    enhanced migratory responses towards chemokines which implied an additional role for SP in promoting leukocyte activation and migration in vivo Mouse macrophages were also used as a model system to elucidate cellular mechanisms of SP-induced chemokine production SP triggered a cascade of molecular events in the cells A key signaling molecule involved was the transcription factor NF-κB SP induced both classical and atypical pathways of NF-κB activation in mouse macrophages Inhibition of NF-κB abolished SP-induced chemokine production by macrophages Furthermore, extracellular signal-regulated kinase (ERK)1/2 lie upstream of NF-κB in SP-induced signal transduction cascade ERK1/2 were activated in the cells between and 30 following SP stimulation A specific ERK1/2 inhibitor prevented activation of NF-κB and chemokine production induced by SP Two kinase pathways, PKCs and PI3K-Akt were further identified to be important for mediating SP-induced ERK1/2 activation PKC isoforms, including the conventional PKCα and novel PKCδ and ε, were selectively activated by SP-NK-1R at early time points (3 min) Inhibition of PKCα, δ, or ε attenuated SP-NK-1R-indcued ERK1/2, NF-κB activation and chemokine production PI3K-Akt pathway was activated at a later time point (15 min) than PKCs and also preceded through ERK1/2 and NF-κB activation to induce chemokine production in macrophages PKCs and PI3K-Akt were two independent pathways and acted synergistically to trigger downstream events PKCs were likely to induce early ERK1/2 activation while PI3K-Akt contributed to the pathway at later time points Collectively, these data indicate that SP-NK-1R triggers two convergent signaling pathways, one led by PKC isoforms and the other by PI3K-Akt, which induce ERK1/2 and NF-κB activation, leading to upregulated chemokine expression in leukocytes vii    Neurokinin A (NKA), another neuropeptide of the tachykinin family, was found to have similar inductive effects on chemokine production in macrophages, mediated via PI3KAkt- and ERK1/2-dependent NF-κB activation viii    LIST OF FIGURES Figure 2.1 CP96,345 treatment attenuated caerulein-induced increase in MCP-1 mRNA expression in the pancreas and lungs Figure 2.2 Effects of caerulein hyperstimulation and CP96,345 treatment on immunohistochemical localization of MCP-1 in the pancreas and lungs Figure 2.3 CP96,345 treatment had no effect on caerulein-induced upregulation of RANTES mRNA expression in the pancreas Figure 2.4 CP96,345 treatment suppressed caerulein-induced upregulation of MIP-1α mRNA expression in the pancreas and lungs Figure 2.5 Effects of caerulein hyperstimulation and CP96,345 treatment on immunohistochemical localization of MIP-1α in the pancreas and lungs Figure 2.6 CP96,345 treatment attenuated caerulein-induced increase in the MIP-2 mRNA expression in the pancreas and lungs Figure 2.7 Effects of caerulein hyperstimulation and CP96,345 treatment on immunohistochemical localization of MIP-2 in the pancreas and lungs Figure 3.1 SP upregulated CD11b integrin (neutrophils activation marker) expression in primary mouse neutrophils Figure 3.2 SP stimulated NF-κB binding activity, IκBα degradation and NF-κB translocation in primary mouse neutrophils Figure 3.3 Effect of SP stimulation on MIP-1α and MIP-2 expression by primary mouse neutrophils Figure 3.4 Effect of SP stimulation on CCR1, CCR2 and CXCR2 expression in primary mouse neutrophils Figure 3.5 SP stimulation enhanced chemotactic responses of primary mouse neutrophils to rMIP-2 and rMIP-1α Figure 4.1 SP enhanced chemokine production in RAW 264.7 macrophages Figure 4.2 SP enhanced chemokine production in primary peritoneal macrophages ix    Figure 4.3 Effect of NK receptor antagonists on SP-induced MIP-2 and MCP-1 upregulation Figure 4.4 SP enhanced NF-κB p65 but not AP-1 DNA-binding activity via the NK-1R Figure 4.5 SP activated NF-κB via the classical and atypical pathways in RAW 264.7 macrophages Figure 4.6 Inhibition of NF-κB activation abolished SP-induced chemokine production in RAW 264.7 cells Figure 4.7 SP induced ERK1/2 activation in RAW 264.7 cells and primary peritoneal macrophages Figure 4.8 Inhibition of ERK1/2 activation attenuated SP-induced chemokine production in RAW 264.7 cells Figure 4.9 SP-induced NF-κB DNA-binding activity was not dependent of ERK1/2 Figure 4.10 ERK1/2 was involved in Ser276-phosphorylation of NF-κB p65 subunit Figure 5.1 SP induced membrane translocation of PKCα, δ, and ε in mouse macrophages Figure 5.2 SP induced phosphorylation of PKCα, δ, and ε in mouse macrophages Figure 5.3 SP-induced PKC translocataion was NK-1R-dependent Figure 5.4 PKCα, δ, and ε were involved in mediating SP-induced MIP-2 production by mouse macrophages Figure 5.5 PKCα, δ, and ε were involved in mediating SP-induced MCP-1 production by mouse macrophages Figure 5.6 PKCα, δ, and ε were involved in mediating SP-induced ERK1/2 activation in mouse macrophages Figure 5.7 PKCα, δ, and ε were involved in mediating SP-induced NF-κB activation in mouse macrophages Figure 5.8 SP induced Akt phosphorylation in mouse macrophages via the NK-1R x    pancreatic acini in response to cerulein hyperstimulation Am J Physiol Gastrointest Liver Physiol 282: G77-G85, 2002b 17 Bhatia M, Brady M, Shokuhi S, Christmas S, Neoptolemos JP, Slavin J Inflammatory mediators in acute pancreatitis J Pathol 190: 117-125, 2000a 18 Bhatia M, Brady M, Zagorski J, Christmas SE, Campbell F, Neoptolemos JP, Slavin J Treatment with neutralising antibody against cytokine induced neutrophil chemoattractant (CINC) protects rats against acute pancreatitis associated lung injury Gut 47: 838-844, 2000b 19 Bhatia M, Wong FL, Cao Y, Lau HY, Huang J, Puneet P, Chevali L.  Pathophysiology of acute pancreatitis. Pancreatology 5: 132-144, 2005 20 Bhatia M, Hofbauer B, Lee HS, Frossard JL, Steer ML The effects of neutrophil depletion on a completely noninvasive model of acute pancreatitis-associated lung injury Int J Pancreatol 24: 77-83, 1998a 21 Bhatia M, Moochhala S Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome J Pathol 202: 145-56, 2004 22 Bhatia M, Neoptolemos JP, Slavin J Inflammatory mediators as therapeutic targets in acute pancreatitis Curr Opin Investig Drugs 2: 496-501, 2001 23 Bhatia M, Ramnath RD, Chevali L, Guglielmotti A Treatment with bindarit, a blocker of MCP-1 synthesis, protects mice against acute pancreatitis Am J Physiol Gastrointest Liver Physiol 288: G1259-G1265, 2005 24 Bhatia M, Saluja AK, Hofbauer B, Frossard JL, Lee HS, Castagliuolo I, Wang CC, Gerard N, Pothoulakis C, Steer ML Role of substance P and the neurokinin receptor in acute pancreatitis and pancreatitis-associated lung injury Proc Natl Acad Sci USA 95: 4760-4765, 1998b 25 Bhatia M, Saluja AK, Hofbauer B, Steer ML Neutral endopeptidase (NEP) plays an anti-inflammatory role in acute pancreatitis and pancreatitis-associated lung injury Pancreas 15: 428, 1997 26 Bhatia M, Sidhapuriwala JN, Wei Ng S, Tamizhselvi R, Moochhala SM.  Proinflammatory effects of hydrogen sulphide on substance P in caerulein-induced acute pancreatitis J Cell Mol Med 12: 580-590, 2008 27 Bhatia M, Slavin J, Cao Y, Basbaum AI, Neoptolemos JP Preprotachykinin-A gene deletion protects mice against acute pancreatitis and associated lung injury Am J Physiol Gastrointest Liver Physiol 284: G830-G836, 2003 28 Bhatia M, Sun J, He M, Hegde A, Ramnath RD Chemokines in acute pancreatitis In: Progress in Chemokine Research, edited by Linkes WP Nova Science Publishers, Inc 2007 29 Bhatia M, Wallig MA, Hofbauer B, Lee HS, Frossard JL, Steer ML, Saluja AK Induction of apoptosis in pancreatic acinar cells reduces the severity of acute pancreatitis Biochem Biophys Res Commun 246: 476-483, 1998c 30 Bhatia M, Wong FL, Cao Y, Lau HY, Huang J, Puneet P, Chevali, L Pathophysiology of acute pancreatitis Pancreatology 5: 132-144, 2005 31 Bhatia M, Zhi L, Zhang H, Ng SW, Moore PK Role of Substance P in hydrogen sulfide-induced pulmonary inflammation in mice Am J Physiol Lung Cell Mol Physiol 291: L896-L904, 2006 178    32 Bilchik AJ, Leach SD, Zucker KA, Modlin IM Experimental models of acute pancreatitis J Surg Res 48: 639-647, 1990 33 Bird TA, Schooley K, Dower SK, Hagen H, Virca GD Activation of nuclear transcription factor NF-kappaB by interleukin-1 is accompanied by casein kinase IImediated phosphorylation of the p65 subunit J Biol Chem 272: 32606-32612, 1997 34 Bless NM, Huber-Lang M, Guo RF, Warner RL, Schmal H, Czermak BJ, Shanley TP, Crouch LD, Lentsch AB, Sarma V, Mulligan MS, Friedl HP, Ward PA Role of CC chemokines (macrophage inflammatory protein-1 beta, monocyte chemoattractant protein-1, RANTES) in acute lung injury in rats J Immunol 164: 2650–2659, 2000 35 Bohuslav J, Chen LF, Kwon H, Mu Y, Greene WC p53 induces NFkappaB activation by an IkappaB kinase-independent mechanism involving phosphorylation of p65 by ribosomal S6 kinase J Biol Chem 279: 26115-26125, 2004 36 Bonecchi R, Polentarutti N, Luini W, Borsatti A, Bernasconi S, Locati M, Power C, Proudfoot A, Wells TN, Mackay C, Mantovani A, Sozzani S Up-regulation of CCR1 and CCR3 and induction of chemotaxis to CC chemokines by IFN-gamma in human neutrophils J Immunol 162: 474–479, 1999 37 Boot JD, de Haas S, Tarasevych S, Roy C, Wang L, Amin D, Cohen J, Sterk PJ, Miller B, Paccaly A, Burggraaf J, Cohen AF, Diamant Z Effect of an NK1/NK2 receptor antagonist on airway responses and inflammation to allergen in asthma Am J Respir Crit Care Med 175: 450-457, 2007 38 Bost KL, Pascual DW Substance P: A late-acting B lymphocyte differentiation cofactor Am J Physiol 262: C537–C545, 1992 39 Bowden JJ, Garland AM, Baluk P, Lefevre P, Grady EF, Vigna SR, Bunnett NW, McDonald DM Direct observation of substance P-induced internalization of neurokinin (NK1) receptors at sites of inflammation Proc Natl Acad Sci USA 91: 8964-8968, 1994 40 Bozic CR, Lu B, Hopken UE, Gerard C, Gerard NP Neurogenic amplification of immune complex inflammation Science 273: 1722-1725, 1996 41 Brady M, Bhatia M, Christmas S, Boyd MT, Neoptolemos JP, Slavin J Expression of the chemokines MCP-1/JE and cytokine-induced neutrophil chemoattractant in early acute pancreatitis Pancreas 25: 260-269, 2002 42 Bremer AA, Leeman SE, Boyd ND The common C-terminal sequences of substance P and neurokinin A contact the same region of the NK-1 receptor FEBS Lett 486: 43-48, 2000 43 Bremer AA, Tansky MF, Wu M, Boyd ND, Leeman SE Direct evidence for the interaction of neurokinin A with the tachykinin NK(1) receptor in tissue Eur J Pharmacol 423: 143-147, 2001 44 Brogden KA, Guthmiller JM, Salzet M, Zasloff M The nervous system and innate immunity: the neuropeptide connection Nat Immunol 6:558-564, 2005 45 Brose N, Rosenmund C Move over protein kinase C, you've got company: alternative cellular effectors of diacylglycerol and phorbol esters J Cell Sci 115: 4399-4411, 2002 46 Brunelleschi S, Nicali R, Lavagno L, Viano I, Pozzi E, Gagliardi L, Ghio P, Albera C Tachykinin activation of human monocytes from patients with interstitial lung disease, healthy smokers or healthy volunteers Neuropeptides 34: 45-50, 2000 179    47 Burcher E, Shang F, Warner FJ, Du Q, Lubowski DZ, King DW, Liu L Tachykinin NK2 receptor and functional mechanisms in human colon: changes with indomethacin and in diverticular disease and ulcerative colitis J Pharmacol Exp Ther 324: 170-8, 2008 48 Cao T, Pinter E, Al-Rashed S, Gerard N, Hoult JR, Brain SD Neurokinin-1 receptor agonists are involved in mediating neutrophil accumulation in the inflamed, but not normal, cutaneous microvasculature: an in vivo study using neurokinin-1 receptor knockout mice J Immunol 164: 5424-5429, 2000 49 Cao Y, Adhikari S, Clement MV, Wallig M, Bhatia M Induction of apoptosis by crambene protects mice against acute pancreatitis via anti-inflammatory pathways Am J Pathol 170: 1521-1534, 2007 50 Carter AB, Knudtson KL, Monick MM, Hunninghake GW The p38 mitogenactivated protein kinase is required for NF-kappaB-dependent gene expression The role of TATA-binding protein (TBP) J Biol Chem 274: 30858-30863, 1999 51 Castagliuolo I, Riegler M, Pasha A, Nikulasson S, Lu B, Gerard C, Gerard NP, Pothoulakis C Neurokinin-1 (NK-1) receptor is required in Clostridium difficileinduced enteritis J Clin Investig 101: 1547–1550, 1998 52 Chang L, Karin M Mammalian MAP kinase signalling cascades Nature 410: 3740, 2001 53 Chen L, Fischle W, Verdin E, Greene WC Duration of nuclear NF-kappaB action regulated by reversible acetylation Science 293: 1653-1657, 2001 54 Chen L, Hahn H, Wu G, Chen CH, Liron T, Schechtman D, Cavallaro G, Banci L, Guo Y, Bolli R, Dorn GW 2nd, Mochly-Rosen D Opposing cardioprotective actions and parallel hypertrophic effects of delta PKC and epsilon PKC Proc Natl Acad Sci U S A 98: 11114-11119, 2001 55 Chen LF, Greene WC Regulation of distinct biological activities of the NFkappaB transcription factor complex by acetylation J Mol Med 81: 549-557, 2003 56 Cheng SS, Lai JJ, Lukacs NW, Kunkel SL Granulocyte-macrophage colony stimulating factor up-regulates CCR1 in human neutrophils J Immunol 166: 1178– 1184, 2001 57 Ciesielski CJ, Andreakos E, Foxwell BM, Feldmann M TNFalpha-induced macrophage chemokine secretion is more dependent on NF-kappaB expression than lipopolysaccharides-induced macrophage chemokine secretion Eur J Immunol 32: 2037-2045, 2002 58 Conti P, Barbacane RC, Di Gioacchino M, Reale M Will MCP-1 and RANTES take center stage in inflammatory diseases including asthma? Allergy Asthma Proc 19: 121-123, 1998 59 Crimi N, Pagano C, Palermo F, Mastruzzo C, Prosperini G, Pistorio MP, Vancheri C Inhibitory effect of a leukotriene receptor antagonist (montelukast) on neurokinin A-induced bronchoconstriction J Allergy Clin Immunol 111: 833-839, 2003 60 Cross LJ, Heaney LG, Ennis M Histamine release from human bronchoalveolar lavage mast cells by neurokinin A and bradykinin Inflamm Res 46: 306-309, 1997 180    61 Deak M, Clifton AD, Lucocq LM, Alessi DR Mitogen- and stress-activated protein kinase-1 (MSK1) is directly activated by MAPK and SAPK2/p38, and may mediate activation of CREB EMBO J 17: 4426-4441, 1998 62 Dempsey EC, Newton AC, Mochly-Rosen D, Fields AP, Reyland ME, Insel PA, Messing RO Protein kinase C isozymes and the regulation of diverse cell responses Am J Physiol Lung Cell Mol Physiol 279: L429-L438, 2000 63 DeRose V, Robbins RA, Snider RM, Spurzem JR, Thiele GM, Rennard SI, Rubinstein I Substance P increases neutrophil adhesion to bronchial epithelial cells J Immunol 152: 1339-1346, 1994 64 Dianzani C, Lombardi G, Collino M, Ferrara C, Cassone MC, Fantozzi R Priming effects of substance P on calcium changes evoked by interleukin-8 in human neutrophils J Leukoc Biol 69: 1013-1018, 2001 65 Dong C, Davis RJ, Flavell RA MAP kinases in the immune response Annu Rev Immunol 20: 55-72, 2002 66 Downward J Mechanisms and consequences of activation of protein kinase B/Akt Curr Opin Cell Biol 10: 262-267, 1998 67 Elbim C, Prevot MH, Bouscarat F, Franzini E, Chollet-Martin S, Hakim J, neutrophils from human Gougerot-Pocidalo MA.  Polymorphonuclear immunodeficiency virus-infected patients show enhanced activation, diminished fMLP-induced L-selectin shedding, and an impaired oxidative burst after cytokine priming. Blood 84: 2759-2766, 1994 68 Fiebich BL, Schleicher S, Butcher RD, Craig A, Lieb K The neuropeptide substance P activates p38 mitogen-activated protein kinase resulting in IL-6 expression independently from NF-kappa B J Immunol 165: 5606–5611, 2000 69 Fink GW, Norman JG Intrapancreatic interleukin-1beta gene expression by specific leukocyte populations during acute pancreatitis.  J Surg Res 63: 369-373, 1996 70 Genovese T, Mazzon E, Di Paola R, Muia C, Crisafulli C, Malleo G, Esposito E, Cuzzocrea S Role of peroxisome proliferator-activated receptor-alpha in acute pancreatitis induced by cerulein Immunology 118: 559-570, 2006 71 Gerard C, Frossard JL, Bhatia M, Saluja A, Gerard NP, Lu B, Steer M Targeted disruption of the beta-chemokine receptor CCR1 protects against pancreatitisassociated lung injury J Clin Invest 100: 2022-2027, 1997 72 Gerard C, Rollins BJ Chemokines and disease Nat Immunol 2: 108-115, 2001 73 Gerritsen ME, Williams AJ, Neish AS, Moore S, Shi Y, Collins T CREB-binding protein/p300 are transcriptional coactivators of p65 Proc Natl Acad Sci U S A 94: 2927-2932, 1997 74 Gharbi SI, Zvelebil MJ, Shuttleworth SJ, Hancox T, Saghir N, Timms JF, Waterfield MD Exploring the specificity of the PI3K family inhibitor LY294002 Biochem J 404: 15-21, 2007 75 Ghosh S, Karin M Missing pieces in the NF-kappaB puzzle Cell 109: S81-S96, 2002 76 Gorelick FS, Otani T Mechanisms of intracellular zymogen activation Baillieres Best Pract Res Clin Gastroenterol 13: 227-240, 1999 181    77 Grady T, Liang P, Ernst SA, Logsdon CD Chemokine gene expression in rat pancreatic acinar cells is an early event associated with acute pancreatitis Gastroenterology 113: 1966-1975, 1997 78 Grady T, Mah'Moud M, Otani T, Rhee S, Lerch MM, Gorelick FS Zymogen proteolysis within the pancreatic acinar cell is associated with cellular injury Am J Physiol 275: G1010-G1017, 1998 79 Grimsholm O, Guo Y, Ny T, Rantapaa-Dahlqvist S, Forsgren S Are neuropeptides important in arthritis? Studies on the importance of bombesin/GRP and substance P in a murine arthritis model Ann N Y Acad Sci 1110: 525-538, 2007 80 Guha M, Mackman N The phosphatidylinositol 3-kinase-Akt pathway limits lipopolysaccharide activation of signaling pathways and expression of inflammatory mediators in human monocytic cells J Biol Chem 277: 32124-32132, 2002 81 Guo L, Yamaguchi Y, Ikei S, Sugita H, Ogawa M Neutrophil elastase inhibitor (ONO-5046) prevents lung hemorrhage induced by lipopolysaccharide in rat model of cerulein pancreatitis Dig Dis Sci 40: 2177-2183, 1995 82 Gupta M, Mahanty S, Ahmed R, Rollin PE Monocyte-derived human macrophages and peripheral blood mononuclear cells infected with ebola virus secrete MIP-1alpha and TNF-alpha and inhibit poly-IC-induced IFN-alpha in vitro Virology 284: 20-25, 2001 83 Haines KA, Kolasinski SL, Cronstein BN, Reibman J, Gold LI, Weissmann G Chemoattraction of neutrophils by substance P and transforming growth factor-beta is inadequately explained by current models of lipid remodeling J Immunol 151: 1491–1499, 1993 84 Harrison S, Geppetti P Substance p Int J Biochem Cell Biol 33: 555-576, 2001 85 Hartwig W, Werner J, Warshaw AL, Antoniu B, Castillo CF, Gebhard MM, Uhl W, Buchler MW Membrane-bound ICAM-1 is upregulated by trypsin and contributes to leukocyte migration in acute pancreatitis Am J Physiol Gastrointest Liver Physiol 287: G1194-1199, 2004 86 Hastrup H, Schwartz TW Septide and neurokinin A are high-affinity ligands on the NK-1 receptor: evidence from homologous versus heterologous binding analysis FEBS Lett 399: 264-266, 1996 87 Hazzalin CA, Mahadevan LC MAPK-regulated transcription: a continuously variable gene switch? Nat Rev Mol Cell Biol 3: 30-40, 2002 88 Hegde A, Zhang H, Moochhala SM, Bhatia M Neurokinin-1 receptor antagonist treatment protects mice against lung injury in polymicrobial sepsis J Leukoc Biol 82: 678-685, 2007 89 Hirsch E, Katanaev VL, Garlanda C, Azzolino O, Pirola L, Silengo L, Sozzani S, Mantovani A, Altruda F, Wymann MP Central role for G protein-coupled phosphoinositide 3-kinase gamma in inflammation Science 287: 1049-1053, 2000 90 Ho WZ, Kaufman D, Uvaydova M, Douglas SD Substance P augments interleukin-10 and tumor necrosis factor-alpha release by human cord blood monocytes and macrophages J Neuroimmunol 71: 73-80, 1996 91 Holst B, Hastrup H, Raffetseder U, Martini L, Schwartz TW Two active molecular phenotypes of the tachykinin NK1 receptor revealed by G-protein fusions and mutagenesis J Biol Chem 276: 19793-19799, 2001 182    92 Horuk R Molecular properties of the chemokine receptor family TIPS 15: 159165, 1994 93 Inoue S, Nakao A, Kishimoto W, Murakami H, Harada A, Nonami T, Takagi H.  LFA-1 (CD11a/CD18) and ICAM-1 (CD54) antibodies attenuate superoxide anion release from polymorphonuclear leukocytes in rats with experimental acute pancreatitis. Pancreas 12: 183-188, 1996 94 Inoue S, Nakao A, Kishimoto W, Murakami H, Itoh K, Itoh T, Harada A, Nonami T, Takagi H Anti-neutrophil antibody attenuates the severity of acute lung injury in rats with experimental acute pancreatitis. Arch Surg 130: 93-98, 1995 95 Isenmann R, Rau B, Beger HG Baterial infection and extent of necrosis are determinants of organ failure in patients with acute necrotising pancreatitis Br J Surg 86: 1020-1024, 1999 96 Jahnz-Rozyk KM, Kuna P, Pirozynska E Monocyte chemotactic and activating factor/monocyte chemoattractant protein (MCAF/MCP-1) in bronchoalveolar lavage fluid from patients with atopic asthma and chronic bronchitis J Investig Allergol Clin Immunol 7: 254-259, 1997 97 Jang BC, Kim DH, Park JW, Kwon TK, Kim SP, Song DK, Park JG, Bae JH, Mun KC, Baek WK, Suh MH, Hla T, Suh SI Induction of cyclooxygenase-2 in macrophages by catalase: role of NF-kappaB and PI3K signaling pathways Biochem Biophys Res Commun 316: 398-406, 2004 98 Jang BC, Paik JH, Kim SP, Shin DH, Song DK, Park JG, Suh MH, Park JW, Suh SI Catalase induced expression of inflammatory mediators via activation of NFkappaB, PI3K/AKT, p70S6K, and JNKs in BV2 microglia Cell Signal 17: 625-633, 2005 99 Johnston B, Burns AR, Suematsu M, Issekutz TB, Woodman RC, Kubes P Chronic inflammation upregulates chemokine receptors and induces neutrophil migration to monocyte chemoattractant protein-1 J Clin Invest 103: 1269–1276, 1999 100 Kao SJ, Lei HC, Kuo CT, Chang MS, Chen BC, Chang YC, Chiu WT, Lin CH Lipoteichoic acid induces nuclear factor-kappaB activation and nitric oxide synthase expression via phosphatidylinositol 3-kinase, Akt, and p38 MAPK in RAW 264.7 macrophages Immunology 115: 366-374, 2005 101 Karimi K, Kool M, Nijkamp FP, and Redegeld FA Substance P can stimulate prostaglandin D2 and leukotriene C4 generation without granule exocytosis in murine mast cells Eur J Pharmacol 489: 49-54, 2004 102 Karin M, Ben-Neriah Y Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity Annu Rev Immunol 18: 621-663, 2000 103 Kefaloyianni E, Gaitanaki C, Beis I ERK1/2 and p38-MAPK signalling pathways, through MSK1, are involved in NF-kappaB transactivation during oxidative stress in skeletal myoblasts Cell Signal 18: 2238-2251, 2006 104 Kiernan R, Bres V, Ng RW, Coudart MP, El Messaoudi S, Sardet C, Jin DY, Emiliani S, Benkirane M Post-activation turn-off of NF-kappa B-dependent transcription is regulated by acetylation of p65 J Biol Chem 278: 2758-2766, 2003 183    105 Killingsworth CR, Shore SA, Alessandrini F, Dey RD, Paulauskis JD Rat alveolar macrophages express preprotachykinin gene-I mRNA-encoding tachykinins Am J Physiol 273: L1073–L1081, 1997 106 Kledal TN, Rosenkilde MM, Coulin F, Simmons G, Johnsen AH, Alouani S, Power CA, Lüttichau HR, Gerstoft J, Clapham PR, Clark-Lewis I, Wells TN, Schwartz TW A broad-spectrum chemokine antagonist encoded by Kaposi's sarcoma-associated herpesvirus Science 277: 1656-1659, 1997 107 Kobayashi M, Takahashi H, Sanford AP, Herndon DN, Pollard RB, Suzuki F.  An increase in the susceptibility of burned patients to infectious complications due to impaired production of macrophage inflammatory protein alpha.  J Immunol 169:4460-4466, 2002 108 Kong L, Santiago N, Han TQ, Zhang SD Clinical characteristics and prognostic factors of severe acute pancreatitis World J Gastroenterol 10: 3336-3338, 2004 109 Koon HW, Zhao D, Zhan Y, Simeonidis S, Moyer MP, Pothoulakis C Substance P-stimulated interleukin-8 expression in human colonic epithelial cells involves protein kinase Cdelta activation J Pharmacol Exp Ther 314: 1393-1400, 2005 110 Kraneveld AD, James DE, de Vries A, Nijkamp FP Excitatory non-adrenergicnon-cholinergic neuropeptides: key players in asthma Eur J Pharmacol 405: 113-129, 2000 111 Kwon D, Fuller AC, Palma JP, Choi IH, Kim BS Induction of chemokines in human astrocytes by picornavirus infection requires activation of both AP-1 and NFkappa B Glia 45: 287-296, 2004 112 Kyriakis JM, Avruch J Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation Physiol Rev 81: 807869, 2001 113 Lagathu C, Eustace B, Prot M, Frantz D, Gu Y, Bastard JP, Maachi M, Azoulay S, Briggs M, Caron M, Capeau J Some HIV antiretrovirals increase oxidative stress and alter chemokine, cytokine or adiponectin production in human adipocytes and macrophages Antivir Ther 12: 489-500, 2007 114 Lakshminarayanan V, Drab-Weiss EA, Roebuck KA H2O2 and tumor necrosis factor-alpha induce differential binding of the redox-responsive transcription factors AP-1 and NF-kappaB to the interleukin-8 promoter in endothelial and epithelial cells J Biol Chem 273: 32670-32678, 1998 115 Lampel M, Kern HF.  Acute interstitial pancreatitis in the rat induced by excessive doses of a pancreatic secretagogue.  Virchows Arch A Pathol Anat Histol 373: 97-117, 1977 116 Lau HY, Bhatia M Effect of CP96,345 on the expression of tachykinins and neurokinin receptors in acute pancreatitis J Pathol 208: 364-371, 2006 117 Lau HY, Bhatia M Effect of CP-96,345 on the expression of adhesion molecules in acute pancreatitis in mice Am J Physiol Gastrointest Liver Physiol 292: G1283-1292, 2007 118 Lau HY, Wong FL, Bhatia M A key role of neurokinin receptors in acute pancreatitis and associated lung injury Biochem Biophys Res Commun 327: 509-15, 2005 184    119 Lavagno L, Bordin G, Colangelo D, Viano I, Brunelleschi S Tachykinin activation of human monocytes from patients with rheumatoid arthritis: in vitro and ex-vivo effects of cyclosporin A Neuropeptides 35: 92-99, 2001 120 Leifeld L, Dumoulin FL, Purr I, Janberg K, Trautwein C, Wolff M, Manns MP, Sauerbruch T, Spengler U Early up-regulation of chemokine expression in fulminant hepatic failure J Pathol 199: 335-344, 2003 121 Lloyds D, Hallett MB Activation and priming of the human neutrophil oxidase response by substance P: distinct signal transduction pathways Biochim Biophys Acta 175: 207–213, 1993 122 Lombardi B, Estes LW, Longnecker DS Acute hemorrhagic pancreatitis (massive necrosis) with fat necrosis induced in mice by DL-ethionine fed with a choline-defi cient diet Am J Pathol 79: 465-480, 1975 123 Lotz M, Vaughan JH, Carson DA Effect of neuropeptides on production of inflammatory cytokines by human monocytes Science 241: 1218-1221, 1988 124 Lukacs NW, Hogaboam C, Campbell E, Kunkel SL Chemokines: function, regulation, and alteration of inflammatory responses Chem Immunol 72:102–120, 1999 125 Luyendyk JP, Schabbauer GA, Tencati M, Holscher T, Pawlinski R, Mackman N Genetic analysis of the role of the PI3K-Akt pathway in lipopolysaccharideinduced cytokine and tissue factor gene expression in monocytes/macrophages J Immunol 180: 4218-4226, 2008 126 Luster AD.  Chemokines chemotactic cytokines that mediate inflammation.  N Engl J Med 338:436-445, 1998 127 Maa J, Grady EF, Yoshimi SK, Drasin TE, Kim EH, Hutter MM, Bunnett NW, Kirkwood KS Substance P is a determinant of lethality in diet-induced hemorrhagic pancreatitis in mice Surgery.128: 232-9, 2000 128 Madrid LV, Mayo MW, Reuther JY, Baldwin AS Jr Akt stimulates the transactivation potential of the RelA/p65 Subunit of NF-kappa B through utilization of the Ikappa B kinase and activation of the mitogen-activated protein kinase p38 J Biol Chem 276: 18934-18940, 2001 129 Madrid LV, Wang CY, Guttridge DC, Schottelius AJ, Baldwin AS Jr, Mayo MW Akt suppresses apoptosis by stimulating the transactivation potential of the RelA/p65 subunit of NF-κB Mol Cell Biol 20: 1626–1638, 2000 130 Maggi CA The mammalian tachykinin receptors Gen Pharmacol 26: 911-944, 1995 131 Maggi CA The effects of tachykinins on inflammatory and immune cells Regul Pept 70: 75-90, 1997 132 Makhija R, Kingsnorth AN Cytokine storm in acute pancreatitis J Hepatobiliary Pancreat Surg 9: 401-410, 2002 133 Marra F Renaming cytokines: MCP-1, major chemokine in pancreatitis Gut 54: 1679-1681, 2005 134 Marriott I, Bost KL IL-4 and IFN-gamma up-regulate substance P receptor expression in murine peritoneal macrophages J Immunol 165: 182-191, 2000 135 Martin AG, San-Antonio B, Fresno M Regulation of nuclear factor kappa B transactivation Implication of phosphatidylinositol 3-kinase and protein kinase C zeta 185    in c-Rel activation by tumor necrosis factor alpha J Biol Chem 276: 15840-15849, 2001 136 Merika M, Williams AJ, Chen G, Collins T, Thanos D Recruitment of CBP/p300 by the IFN beta enhanceosome is required for synergistic activation of transcription Mol Cell 1: 277-287, 1998 137 Metwali A, Blum AM, Elliott DE, Setiawan T, Weinstock JV Cutting edge: hemokinin has substance P-like function and expression in inflammation J Immunol 172: 6528-6532, 2004 138 McKinley L, Kim J, Bolgos GL, Siddiqui J, Remick DG CXC chemokines modulate IgE secretion and pulmonary inflammation in a model of allergic asthma Cytokine 32: 178-185, 2005 139 Mikami Y, Takeda K, Shibuya K, Qiu-Feng H, Shimamura H, Yamauchi J, Egawa S, Sunamura M, Yagi H, Endo Y, Matsuno S.  Do peritoneal macrophages play an essential role in the progression of acute pancreatitis in rats? Pancreas 27: 253-260, 2003 140 Mookerjee Basu J, Mookerjee A, Sen P, Bhaumik S, Banerjee S, Naskar K, Choudhuri SK, Saha B, Raha S, Roy S Sodium antimony gluconate induces generation of reactive oxygen species and nitric oxide via phosphoinositide 3-kinase and mitogen-activated protein kinase activation in Leishmania donovani-infected macrophages Antimicrob Agents Chemother 50: 1788-1797, 2006 141 Moynagh PN The NF-kappaB pathway J Cell Sci 118: 4589-4592, 2005 142 Murakami H, Nakao A, Kishimoto W, Nakano M, Takagi H Detection of O2generation and neutrophil accumulation in rat lungs after acute necrotizing pancreatitis Surgery 118: 547-554, 1995 143 Murphy PM The molecular biology of leukocyte chemoattractant receptors Annu Rev Immunol 12: 593-633, 1994 144 Nakamichi I, Habtezion A, Zhong B, Contag CH, Butcher EC, Omary MB Hemin-activated macrophages home to the pancreas and protect from acute pancreatitis via heme oxygenase-1 induction J Clin Invest 115: 3007-3014, 2005 145 Naumann M, Scheidereit C Activation of NF-kappa B in vivo is regulated by multiple phosphorylations EMBO J 13:4597-4607, 1994 146 Ng SW, Zhang H, Hegde A, Bhatia M Role of preprotachykinin-A gene products on multiple organ injury in LPS-induced endotoxemia J Leukoc Biol 83: 288-295, 2008 147 Noble MD, Romac J, Wang Y, Hsu J, Humphrey JE, Liddle RA Local disruption of the celiac ganglion inhibits substance P release and ameliorates caerulein-induced pancreatitis in rats Am J Physiol Gastrointest Liver Physiol 291, G128-G134, 2006 148 O'Connor TM, O'Connell J, O'Brien DI, Goode T, Bredin CP, Shanahan F The role of substance P in inflammatory disease J Cell Physiol 201: 167-80, 2004 149 Ohno S, Nishizuka Y Protein kinase C isotypes and their specific functions: prologue J Biochem 132: 509-511, 2002 150 Ozes ON, Mayo LD, Gustin JA, Pfeffer SR, Pfeffer LM, Donner DB NFkappaB activation by tumour necrosis factor requires the Akt serine-threonine kinase Nature 401: 82-85, 1999 186    151 Page NM Hemokinins and endokinins Cell Mol Life Sci 61: 1652–1663, 2004 152 Palma C Tachykinins and their receptors in human malignancies Curr Drug Targets 7: 1043-1052, 2006 153 Parekh DB, Ziegler W, Parker PJ Multiple pathways control protein kinase C phosphorylation EMBO J 19: 496-503, 2000 154 Pastor CM, Rubbia-Brandt L, Hadengue A, Jordan M, Morel P, Frossard JL Role of macrophage inflammatory peptide-2 in cerulein-induced acute pancreatitis and pancreatitis-associated lung injury Lab Invest 83: 471-478, 2003 155 Pastor CM, Vonlaufen A, Georgi F, Hadengue A, Morel P, Frossard JL Neutrophil depletion but not prevention of Kupffer cell activation decreases the severity of cerulein-induced acute pancreatitis World J Gastroenterol 12: 1219-1224, 2006 156 Patel DN, King CA, Bailey SR, Holt JW, Venkatachalam K, Agrawal A, Valente AJ, Chandrasekar B Interleukin-17 stimulates C-reactive protein expression in hepatocytes and smooth muscle cells via p38 MAPK and ERK1/2-dependent NFkappaB and C/EBPbeta activation J Biol Chem 282: 27229-27238, 2007 157 Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K, Cobb MH Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions Endocr Rev 22: 153-183, 2001 158 Pengal RA, Ganesan LP, Wei G, Fang H, Ostrowski MC, Tridandapani S Lipopolysaccharide-induced production of interleukin-10 is promoted by the serine/threonine kinase Akt Mol Immunol 43: 1557-1564, 2006 159 Pernow B Substance P Pharmacol Rev 35: 85–141, 1983 160 Pierce JW, Schoenleber R, Jesmok G, Best J, Moore SA, Collins T, Gerritsen ME Novel inhibitors of cytokine-induced IkappaBalpha phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects in vivo J Biol Chem 272: 21096-21103, 1997 161 Pinter E, Brown B, Hoult JR, Brain SD Lack of evidence for tachykinin NK1 receptor-mediated neutrophil accumulation in the rat cutaneous microvasculature by thermal injury Eur J Pharmacol 369: 91-98, 1999 162 Pischke SE, Zhou Z, Song R, Ning W, Alam J, Ryter SW, Choi AM Phosphatidylinositol 3-kinase/Akt pathway mediates heme oxygenase-1 regulation by lipopolysaccharide Cell Mol Biol (Noisy-le-grand) 51: 461-470, 2005 163 Poch B, Gansauge F, Rau B, Wittel U, Gansauge S, Nüssler AK, Schoenberg M, Beger HG The role of polymorphonuclear leukocytes and oxygen-derived free radicals in experimental acute pancreatitis: mediators of local destruction and activators of inflammation FEBS Lett 461: 268-72, 1999 164 Puneet P, Hegde A, Ng SW, Lau HY, Lu J, Moochhala SM, Bhatia M Preprotachykinin-A gene products are key mediators of lung injury in polymicrobial sepsis J Immunol 176: 3813-3820, 2006 165 Ramnath RD, Bhatia M Substance P treatment stimulates chemokine synthesis in pancreatic acinar cells via the activation of NF-kappaB Am J Physiol Gastrointest Liver Physiol 291: G1113-G1119, 2006 187    166 Ramnath RD, Sun J, Adhikari S, Zhi L, Bhatia M Role of PKC-delta on substance P-induced chemokine synthesis in pancreatic acinar cells Am J Physiol Cell Physiol 294: C683-C692, 2008 167 Rau B, Baumgart K, Kruger CM, Schilling M, Beger HG CC-chemokine activation in acute pancreatitis: enhanced release of monocyte chemoattractant protein-1 in patients with local and systemic complications Intensive Care Med 29: 622-629, 2003 168 Regoli D, Rhaleb NE, Dion S, Tousignant C, Rouissi N, Jukic D, Drapeau G Neurokinin A A pharmacological study Pharmacol Res 22: 1-14, 1990 169 Roebuck KA, Carpenter LR, Lakshminarayanan V, Page SM, Moy JN, Thomas LL Stimulus-specific regulation of chemokine expression involves differential activation of the redox-responsive transcription factors AP-1 and NF-kappaB J Leukoc Biol 65: 291-298, 1999 170 Rollins BJ Chemokines Blood 90: 909-928, 1999 171 Roux PP, Blenis J ERK and p38 MAPK-activated protein kinases: a family of protein kinases with diverse biological functions Microbiol Mol Biol Rev 68: 320344, 2004 172 Saban MR, Saban R, Bjorlling D, Haak-Fendscho M Involvement of leukotrienes, TNF-alpha, and the LFA-1/ICAM-1 interaction in substance P-induced granulocyte inflitration J Leukoc Biol 61: 445-451, 1997 173 Sagan S, Chassaing G, Pradier L, Lavielle S Tachykinin peptides affect differently the second messenger pathways after binding to CHO-expressed human NK-1 receptors J Pharmacol Exp Ther 276: 1039-1048, 1996 174 Sandoval D, Gukovskaya A, Reavey P, Gukovsky S, Sisk A, Braquet P, Pandol SJ, Poucell-Hatton S.  The role of neutrophils and platelet-activating factor in mediating experimental pancreatitis. Gastroenterology 111: 1081-1091, 1996 175 Sapna S, Shivakumar K Substance P enhances soluble ICAM-1 release from adult rat cardiac fibroblasts by a p42/44 MAPK- and PKC-mediated mechanism Cell Biol Int 31: 856-859, 2007 176 Schafer C, Tietz AB, Goke B Pathophysiology of acute experimental pancreatitis: lessons from genetically engineered animal models and new molecular approaches Digestion 71: 162-172, 2005 177 Schechtman D, Mochly-Rosen D Isozyme-specific inhibitors and activators of protein kinase C Methods Enzymol 345: 470-489, 2002 178 Scheele GA, Palade GE Studies on the guinea pig pancreas Parallel discharge of exocrine enzyme activities J Biol Chem 250: 2660-2670, 1975 179 Sebastian C, Serra M, Yeramian A, Serrat N, Lloberas J, Celada A Deacetylase activity is required for STAT5-dependent GM-CSF functional activity in macrophages and differentiation to dendritic cells J Immunol 180: 5898-5906, 2008 180 Senninger N, Moody FG, Coelho JC, Van Buren DH The role of biliary obstruction in the pathogenesis of acute pancreatitis in the opossum Surgery 99: 688-693, 1986 181 Serra MC, Bazzoni F, Della Bianca V, Greskowiak M, Rossi F Activation of human neutrophils by substance P Effect on oxidative metabolism, exocytosis, 188    cytosolic Ca2+ concentration and inositol phosphate formation J Immunol 141: 2118–2124, 1998 182 Serra MC, Calzetti F, Ceska V, Cassatella MA Effect of substance P on superoxide anion and IL-8 production by human PMNL Immunology 82: 63–69, 1994 183 Severini C, Improta G, Falconieri-Erspamer G, Salvadori S, Erspamer V The tachykinin peptide family Pharmacol Rev 54: 285-322, 2002 184 Sheppard KA, Rose DW, Haque ZK, Kurokawa R, McInerney E, Westin S, Thanos D, Rosenfeld MG, Glass CK, Collins T Transcriptional activation by NFkappaB requires multiple coactivators Mol Cell Biol 19: 6367-6378, 1999 185 Sinclair MT, McCarthy A, McKay C, Sharples CE, Imrie CW (Abstract) The increasing incidence and high mortality rate from acute pancreatitis in Scotland over the last ten years Gastroenterology 112: A482, 1997 186 Sizemore N, Leung S, Stark GR Activation of phosphatidylinositol 3-kinase in response to interleukin-1 leads to phosphorylation and activation of the NF-kappaB p65/RelA subunit Mol Cell Biol 19:4798-4805, 1999 187 Smiley ST, King JA, Hancock WW Fibrinogen stimulates macrophage chemokine secretion through toll-like receptor J Immunol 167: 2887-2894, 2001 188 Smith MV, Lee MJ, Islam AS, Rohrer JL, Goldberg VM, Beidelschies MA, Greenfield EM Inhibition of the PI3K-Akt signaling pathway reduces tumor necrosis factor-alpha production in response to titanium particles in vitro J Bone Joint Surg Am 89: 1019-1027, 2007 189 Soltoff SP Rottlerin: an inappropriate and ineffective inhibitor of PKCdelta Trends Pharmacol Sci 28: 453-458, 2007 190 Speyer CL, Gao H, Rancilio NJ, Neff TA, Huffnagle GB, Sarma JV, Ward PA Novel chemokine responsiveness and mobilization of neutrophils during sepsis Am J Pathol 165: 2187-2196, 2004 191 Sterner-Kock A, Braun R, van der Vliet A, Schrenzel M, McDonald R, Kabbur M, Vulliet R, Hyde D Substance P primes the formation of hydrogen peroxide and nitric oxide in human neutrophils J Leukoc Biol 65: 834-840, 1999 192 Steer ML.  Relationship between pancreatitis and lung diseases.  Respir Physiol 128:13-6, 2001 193 Sun J, Bhatia M Blockade of neurokinin-1 receptor attenuates CC and CXC chemokine production in experimental acute pancreatitis and associated lung injury Am J Physiol Gastrointest Liver Physiol 292: G143-G153, 2007 194 Sun J, Ramnath RD, Bhatia M Neuropeptide substance P upregulates chemokine and chemokine receptor expression in primary mouse neutrophils Am J Physiol Cell Physiol 293: C696-C704, 2007 195 Sun J, Ramnath RD, Zhi L, Tamizhselvi R, Bhatia M Substance P enhances NF{kappa}B transactivation and chemokine response in murine macrophages via ERK1/2 and p38 MAPK signaling pathways Am J Physiol Cell Physiol In press, 2008 196 Susan D Brain, Helen M Cox Neuropeptides and their receptors: innovative science providing novel therapeutic targets Br J Pharmacol 147: S202–S211, 2006 189    197 Talamini G, Bassi C, Falconi M, Sartori N, Frulloni L, Di Francesco V, Vesentini S, Pederzoli P, Cavallini G Risk of death from acute pancreatitis Role of early, simple "routine" data Int J Pancreatol 19: 15-24, 1996 198 Tamizhselvi R, Moore PK, Bhatia M.  Hydrogen sulfide acts as a mediator of inflammation in acute pancreatitis: in vitro studies using isolated mouse pancreatic acinar cells J Cell Mol Med 11: 315-326, 2007 199 Thrower EC, Osgood S, Shugrue CA, Kolodecik TR, Chaudhuri AM, Reeve Jr JR, Pandol SJ, Gorelick FS The novel protein kinase C isoforms-{delta} and {epsilon} modulate caerulein-induced zymogen activation in pancreatic acinar cells Am J Physiol Gastrointest Liver Physiol In press, 2008 200 Thurston G, Baluk P, Hirata A, McDonald DM Permeability-related changes revealed at endothelial cell borders in inflamed venules by lectin binding Am J Physiol 271: H2547-H2562, 1996 201 Tokuda M, Miyamoto R, Sakuta T, Nagaoka S, Torii M Substance P activates p38 mitogen-activated protein kinase to promote IL-6 induction in human dental pulp fibroblasts Connect Tissue Res 46: 153-158, 2005 202 Trafton JA, Abbadie C, Basbaum AI Differential contribution of substance P and neurokinin A to spinal cord neurokinin-1 receptor signaling in the rat J Neurosci 21: 3656-3664, 2001 203 Trushin SA, Pennington KN, Carmona EM, Asin S, Savoy DN, Billadeau DD, Paya CV Protein kinase Calpha (PKCalpha) acts upstream of PKCtheta to activate IkappaB kinase and NF-kappaB in T lymphocytes Mol Cell Biol 23: 7068-7081, 2003 204 Tsukamoto K, Hazeki K, Hoshi M, Nigorikawa K, Inoue N, Sasaki T, Hazeki O Critical roles of the p110 beta subtype of phosphoinositide 3-kinase in lipopolysaccharide-induced Akt activation and negative regulation of nitrite production in RAW 264.7 cells J Immunol 180: 2054-2061, 2008 205 Verdegaal EM, Zegveld ST, Blokland I, Beekhuizen H, Bakker W, Willems LN, van Furth R Expression of adhesion molecules on granulocytes and monocytes from patients with asthma stimulated in vitro with interleukin-8 and monocyte chemotactic protein-1 Inflammation 22: 229-242, 1998 206 Vermeulen L, De Wilde G, Van Damme P, Vanden Berghe W, Haegeman G Transcriptional activation of the NF-kappaB p65 subunit by mitogen- and stressactivated protein kinase-1 (MSK1) EMBO J 22: 1313-1324, 2003 207 Viatour P, Merville MP, Bours V, Chariot A Phosphorylation of NF-kappaB and IkappaB proteins: implications in cancer and inflammation Trends Biochem Sci 30: 43-52, 2005 208 Ward SG, Finan P Isoform-specific phosphoinositide 3-kinase inhibitors as therapeutic agents Curr Opin Pharmacol 3: 426-434, 2003 209 Wells TN, Power CA, Lusti-Narasimhan M, Hoogewerf AJ, Cooke RM, Chung CW, Peitsch MC, Proudfoot AE.  Selectivity and antagonism of chemokine receptors.  J Leukoc Biol 59: 53-60, 1996 210 White JR, Lee JM, Young PR, Hertzberg RP, Jurewicz AJ, Chaikin MA, Widdowson K, Foley JJ, Martin LD, Griswold DE, Sarau HM Identification of a 190    potent, selective non-peptide CXCR2 antagonist that inhibits interleukin-8-induced neutrophil migration J Biol Chem 273: 10095-10098, 1998 211 Wijkhuisen A, Sagot MA, Frobert Y, Creminon C, Grassi J, Boquet D, Couraud JY Identification in the NK1 tachykinin receptor of a domain involved in recognition of neurokinin A and septide but not of substance P FEBS Lett 447: 155-159, 1999 212 Williams R, Zou X, Hoyle GW Tachykinin-1 receptor stimulates proinflammatory gene expression in lung epithelial cells through activation of NFkappaB via a G(q)-dependent pathway Am J Physiol Lung Cell Mol Physiol 292: L430-L437, 2007 213 Wozniak A, Betts WH, McLennan G, Scicchitano R Activation of human neutrophils by tachykinins: effect on formyl-methionyl-leucyl-phenylalanine- and platelet-activating factor-stimulated superoxide anion production and antibodydependent cell-mediated cytotoxicity Immunology 78: 629-634, 1993 214 Wozniak A, McLennan G, Betts WH, Murphy GA, Scicchitano R Activation of human neutrophils by substance P: effect on fMLPstimulated oxidative and arachidonic acid metabolism and on antibodydependent cell-mediated cytotoxicity Immunology 68: 359-364, 1989 215 Yang BM, Demaine AG, Kingsnorth A Chemokines MCP-1 and RANTES in isolated rat pancreatic acinar cells treated with CCK and ethanol in vitro Pancreas 21: 22-31, 2000 216 Yaraee R, Ebtekar M, Ahmadiani A, Sabahi F, Ghazanfari T The effect of substance P on nitric oxide production by HSV-1 infected macrophages Int Immunopharmacol 7: 135-139, 2007 217 Yoo JK, Kwon H, Khil LY, Zhang L, Jun HS, Yoon JW IL-18 induces monocyte chemotactic protein-1 production in macrophages through the phosphatidylinositol 3-kinase/Akt and MEK/ERK1/2 pathways J Immunol 175: 8280-8286, 2005 218 Zampetaki A, Mitsialis SA, Pfeilschifter J, Kourembanas S Hypoxia induces macrophage inflammatory protein-2 (MIP-2) gene expression in murine macrophages via NF-kappaB: the prominent role of p42/ p44 and PI3 kinase pathways FASEB J 18: 1090-1092, 2004 219 Zhang H, Hegde A, Ng SW, Adhikari S, Moochhala SM, Bhatia M Hydrogen sulfide up-regulates substance P in polymicrobial sepsis-associated lung injury J Immunol 179: 4153-4160, 2007 220 Zhang Y, Berger A, Milne CD, Paige CJ Tachykinins in the immune system Curr Drug Targets 7: 1011-1020, 2006 221 Zhao D, Kuhnt-Moore S, Zeng H, Pan A, Wu JS, Simeonidis S, Moyer MP, Pothoulakis C Substance P-stimulated interleukin-8 expression in human colonic epithelial cells involves Rho family small GTPases Biochem J 368: 665-672, 2002 222 Zagorski J, Wahl SM Inhibition of acute peritoneal inflammation in rats by a cytokine-induced neutrophil chemoattractant receptor antagonist J Immunol 159: 1059-1062, 1997 223 Zhi L, Ang AD, Zhang H, Moore PK, Bhatia M Hydrogen sulfide induces the synthesis of proinflammatory cytokines in human monocyte cell line U937 via the ERK-NF-kappaB pathway J Leukoc Biol 81: 1322-1332, 2007 191    224 Zhong H, May MJ, Jimi E, Ghosh S The phosphorylation status of nuclear NFkappa B determines its association with CBP/p300 or HDAC-1 Mol Cell 9: 625-636, 2002 225 Zhong H, Voll RE, Ghosh S Phosphorylation of NF-kappa B p65 by PKA stimulates transcriptional activity by promoting a novel bivalent interaction with the coactivator CBP/p300 Mol Cell 1: 661-671, 1998 226 Zlotnik A, Yoshie O.  Chemokines: a new classification system and their role in immunity. Immunity 12:121-127, 2000 192    ... infiltrating leukocytes (neutrophils and macrophages) in the pancreas are the main sources of chemokine production in caerulein-induced acute pancreatitis in mice and the alveolar macrophages, epithelial... partially associated with the suppression of SP expression in the pancreas and lungs in the condition (Lau and Bhatia, 2006) Despite the clear pathogenetic role of SP in acute pancreatitis, the. .. acute pancreatitis 10    1.3 SP SP is an 11-amino acid peptide product of the preprotachykinin-A (PPT-A) gene It belongs to the tachykinin family of neuropeptides which also include neurokinin