Activation of Nuclear Factor kappa B and Severe Hepatic Necrosis May Mediate Systemic Inflammation in Choline-deficient/Ethionine-supplemented Diet-induced Pancreatitis

Objectives: We hypothesized that hepatic injury is associated with severe acute pancreatitis (SAP) and may result in lung injury through nuclear factor kappa B (NF-&kgr;B)-dependent inflammatory mediators. The study characterizes the timing and determines the involvement of selected cytokines and chemokines in the pathogenesis of hepatocellular injury associated with SAP. Methods: The SAP was induced in C57BL/6 mice by feeding a choline-deficient/ethionine-supplemented diet. The mice were killed at 12-hour intervals for 96 hours. Terminal deoxynucleotidyl transferase-mediated nick-end labeling staining was used to determine the extent of hepatic apoptosis. The NF-&kgr;B activation in nuclear protein extracts from liver tissue was measured using a sensitive RelA enzyme-linked immunoadsorbent assay. Tumor necrosis factor &agr;, interleukin 6, macrophage inflammatory protein (MIP) 2, and keratinocyte-derived chemokine (KC) levels in homogenates of liver and lung tissues were measured by enzyme-linked immunoadsorbent assay. The SAP-associated neutrophil lung inflammation was measured as tissue myeloperoxidase activity. Results: The SAP and subsequent liver injury were confirmed by histological analysis and rises in plasma amylase and transaminase levels. Severe hepatocellular apoptosis was detected at 36 and 48 hours after the diet initiation by terminal deoxynucleotidyl transferase-mediated nick-end labeling staining (P < 0.05) and subsequently progressed to hepatic necrosis. Liver NF-&kgr;B activation was detected at 36 hours (P < 0.05) and followed by a sharp increase in hepatocellular levels of interleukin 6, MIP-2, and KC at 72 hours and thereafter (P < 0.05). Levels of MIP-2 and KC in lung tissue were also elevated at 72 hours (P < 0.05) and closely correlated with increased myeloperoxidase activity and increased inflammatory cell infiltrate in the lung. Conclusions: Choline-deficient/ethionine-supplemented diet-induced SAP is accompanied with hepatocellular apoptosis and eventual necrosis. This injury is associated with the hepatic NF-&kgr;B activation leading to the production of NF-&kgr;B-dependent cytokines and chemokines in the liver, which may mediate the lung injury.

[1]  Allan R. Brasier,et al.  Identification of an NF-κB-Dependent Gene Network in Cells Infected by Mammalian Reovirus , 2006, Journal of Virology.

[2]  S. Sarna,et al.  Early prediction of organ failure by combined markers in patients with acute pancreatitis , 2005, The British journal of surgery.

[3]  Jun Yang,et al.  Acute pancreatitis induces FasL gene expression and apoptosis in the liver. , 2004, The Journal of surgical research.

[4]  L. Leindler,et al.  Importance of Cytokines, Nitric Oxide, and Apoptosis in the Pathological Process of Necrotizing Pancreatitis in Rats , 2004, Pancreas.

[5]  R. Homer,et al.  Activation of the STAT pathway in acute lung injury. , 2004, American journal of physiology. Lung cellular and molecular physiology.

[6]  Soo Young Lee,et al.  Fas-associated Factor-1 Inhibits Nuclear Factor-κB (NF-κB) Activity by Interfering with Nuclear Translocation of the RelA (p65) Subunit of NF-κB* , 2004, Journal of Biological Chemistry.

[7]  S. Egawa,et al.  Do Peritoneal Macrophages Play an Essential Role in the Progression of Acute Pancreatitis in Rats? , 2003, Pancreas.

[8]  B. Molitoris,et al.  A novel method to determine specificity and sensitivity of the TUNEL reaction in the quantitation of apoptosis. , 2003, American journal of physiology. Cell physiology.

[9]  L. Rubbia‐Brandt,et al.  Role of Macrophage Inflammatory Peptide-2 in Cerulein-Induced Acute Pancreatitis and Pancreatitis-Associated Lung Injury , 2003, Laboratory Investigation.

[10]  W. Chapman,et al.  Systemic nf-?B activation in a transgenic mouse model of acute pancreatitis 1 1 Supported by a VA/HB , 2003 .

[11]  S. Mastorides,et al.  Liver injury during acute pancreatitis: The role of pancreatitis-associated ascitic fluid (paaf), p38-MAPK, and caspase-3 in inducing hepatocyte apoptosis , 2003, Journal of Gastrointestinal Surgery.

[12]  J. Sastre,et al.  Role of cytokines and oxidative stress in the pathophysiology of acute pancreatitis: therapeutical implications. , 2002, Current drug targets. Inflammation and allergy.

[13]  G. Dranoff,et al.  In vivo evidence for the role of GM-CSF as a mediator in acute pancreatitis-associated lung injury. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[14]  K. Reddy,et al.  Acute hepatitis C: To treat or not to treat? , 2002, Hepatology.

[15]  S. Mastorides,et al.  Pancreatic elastase induces liver injury by activating cytokine production within Kupffer cells via nuclear factor-kappa B , 2002, Journal of Gastrointestinal Surgery.

[16]  J. Maher,et al.  Anti-Fas Induces Hepatic Chemokines and Promotes Inflammation by an NF-κB-independent, Caspase-3-dependent Pathway* , 2001, The Journal of Biological Chemistry.

[17]  J. Christman,et al.  Impaired Pulmonary NF-κB Activation in Response to Lipopolysaccharide in NADPH Oxidase-Deficient Mice , 2001, Infection and Immunity.

[18]  S. Pandol,et al.  Localized pancreatic NF-kappaB activation and inflammatory response in taurocholate-induced pancreatitis. , 2001, American journal of physiology. Gastrointestinal and liver physiology.

[19]  A. Masamune,et al.  Ascites of Rat Experimental Model of Severe Acute Pancreatitis Induces Lung Injury , 2001, Pancreas.

[20]  Jun Yang,et al.  Pancreatic elastase activates pulmonary nuclear factor kappa B and inhibitory kappa B, mimicking pancreatitis-associated adult respiratory distress syndrome. , 2000, Surgery.

[21]  Y. Takeyama,et al.  Macrophage-derived transforming growth factor-beta1 induces hepatocellular injury via apoptosis in rat severe acute pancreatitis. , 2000, Surgery.

[22]  Jun Yang,et al.  Elastase mimics pancreatitis-induced hepatic injury via inflammatory mediators. , 2000, The Journal of surgical research.

[23]  W. Chapman,et al.  Hepatic cryoablation, but not radiofrequency ablation, results in lung inflammation. , 2000, Annals of surgery.

[24]  J. Roselló-Catafau,et al.  Activation of alveolar macrophages in lung injury associated with experimental acute pancreatitis is mediated by the liver. , 1999, Annals of surgery.

[25]  H. Reber,et al.  Hepatic Kupffer cell blockade reduces mortality of acute hemorrhagic pancreatitis in mice , 1998, Journal of Gastrointestinal Surgery.

[26]  N. Molan,et al.  [Hepatic lesion in experimental acute pancreatitis. Influence of pancreatic enzyme storage reduction]. , 1998, Revista do Hospital das Clínicas.

[27]  M. Franz,et al.  Tissue-Specific Cytokine Production During Experimental Acute Pancreatitis (A Probable Mechanism for Distant Organ Dysfunction) , 1997, Digestive Diseases and Sciences.

[28]  Jun Yang,et al.  Evidence for an unknown component of pancreatic ascites that induces adult respiratory distress syndrome through an interleukin-1 and tumor necrosis factor-dependent mechanism. , 1997, Surgery.

[29]  R. McIntyre,et al.  Neutrophil depletion attenuates endotoxin-induced dysfunction of cGMP-mediated pulmonary vasorelaxation. , 1996, The American journal of physiology.

[30]  F. Gannon,et al.  A one-hour minipreparation technique for extraction of DNA-binding proteins from animal tissues. , 1994, BioTechniques.

[31]  C. Delaney,et al.  The effect of caerulein induced pancreatitis on the hepatic microvasculature , 1990, The British journal of surgery.

[32]  S. Goldblum,et al.  Lung myeloperoxidase as a measure of pulmonary leukostasis in rabbits. , 1985, Journal of applied physiology.

[33]  J. Ranson,et al.  The Role of Peritoneal Lavage in Severe Acute Pancreatitis , 1978, Annals of surgery.

[34]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[35]  D. Longnecker,et al.  Acute hemorrhagic pancreatitis (massive necrosis) with fat necrosis induced in mice by DL-ethionine fed with a choline-deficient diet. , 1975, The American journal of pathology.

[36]  Soo Young Lee,et al.  Fas-associated factor-1 inhibits nuclear factor-kappaB (NF-kappaB) activity by interfering with nuclear translocation of the RelA (p65) subunit of NF-kappaB. , 2004, The Journal of biological chemistry.

[37]  W. Chapman,et al.  Systemic nf-kappaB activation in a transgenic mouse model of acute pancreatitis. , 2003, The Journal of surgical research.

[38]  Jun Yang,et al.  Regulation of Kupffer cell TNF gene expression during experimental acute pancreatitis: The role of p38-MAPK, ERK1/2, SAPK/JNK, and NF-κB , 2003, Journal of Gastrointestinal Surgery.

[39]  H. Ho,et al.  Hematin is one of the cytotoxic factors in pancreatitis-associated ascitic fluid that causes hepatocellular injury. , 2002, Surgery.

[40]  Y. Takeyama,et al.  Apoptotic cell death of hepatocytes in rat experimental severe acute pancreatitis. , 2000, Surgery.

[41]  A. Lentsch,et al.  Enhanced pulmonary expression of CXC chemokines during hepatic ischemia/reperfusion-induced lung injury in mice. , 1999, The Journal of surgical research.

[42]  M. Melhem,et al.  Acute hemorrhagic pancreatitis in mice: improved survival after indomethacin administration. , 1989, Digestion.

[43]  E. Döpp,et al.  The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3. , 1985, Immunology.

[44]  E. Döpp,et al.  The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in rat recognized by monoclonal antibodies ED1, ED2 and ED3. , 1985, Advances in experimental medicine and biology.

[45]  R. D. Steele.  Role of 3-ethylthiopropionate in ethionine metabolism and toxicity in rats. , 1982, The Journal of nutrition.