1. Volume 122, Issue 2, Pages 448-457 (February 2002)
NF-κB activation in pancreas induces pancreatic and systemic inflammatory response 
Xueqing Chen, Baoan Ji, Bing Han, Stephen A. Ernst, Diane Simeone, Craig D. Logsdon 
Gastroenterology 
Volume 122, Issue 2, Pages (February 2002)
DOI: /gast
Copyright © 2002 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Adenoviral delivery to the pancreas results in acinar cell infection, expression of RelA/p65, and activation of NF-κB. Adp65 adenovirus, which expresses both RelA/p65 and GFP from separate cytomegalovirus promoters, was delivered to rat pancreas in vivo by infusion into the common bile duct. (A) Distribution of viral infection. After 16 hours, portions of the pancreas were processed for histologic analysis. Overlay of GFP fluorescence and the Nomarski image shows the distribution of Adp65 gene transfer. GFP-expressing acinar cells are scattered across the section, appearing singly or, more commonly, in groups within individual acini. Other pancreatic cell types were also infected but at lower numbers, reflecting their relative abundance. (B) Detection of GFP expression in pancreas and lung after adenoviral delivery. Portions of the pancreas and lung were removed 20 hours after adenoviral administration, and equal amounts of protein (20 μg) were analyzed for GFP by Western blotting. (C) Effects of Adp65 on pancreatic levels of RelA/p65. Portions of the pancreas were removed 16 hours after adenoviral administration and analyzed for RelA/p65 by Western blotting. As a protein loading control, the blots were stripped and reprobed for extracellular signal–regulated kinases. (D) NF-κB activation was induced by Adp65 treatment and was partially blocked by coadministration of AdIκB-α. Pancreas from normal rats or rats treated with AdGFP, Adp65, Adp65 plus AdIκB-α for 16 hours or cerulein (10 μg/kg/h intravenously) for 90 minutes were homogenized, nuclear extracts were prepared, and an electrophoretic mobility shift assay was performed. Unlabeled NF-κB oligonucleotide (cold NF-κB) was included to show specificity of DNA binding. HeLa cell nuclear extract was also included as a positive control. (E) Administration of Adp65 increases the level of messenger RNA for the NF-κB–dependent gene, mob-1. Shown is a Northern blot for mob-1 messenger RNA and the ethidium bromide–stained gel as an RNA loading control.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

3. Fig. 2
Administration of Adp65 induces increased MPO activity in the pancreas in a time- and dose-dependent manner, and these effects are inhibited by coadministration of AdIκB-α. (A) Animals were uninjected (none) or injected with control (AdGFP) or NF-κB–activating (Adp65) virus (3 × 107 pfu) and then killed after 16 hours (hatched bars) or 48 hours (open bars), and samples of pancreas were collected. MPO activity was quantitated as an estimation of neutrophilic infiltration. (B) Animals were injected with indicated amounts of control (AdGFP) or NF-κB–activating (Adp65) virus for 16 hours, and the level of pancreatic MPO was determined. (C) Equivalent amounts of viruses of Adp65, AdIκB-α, and AdGFP (3 × 107 pfu) were delivered to rats via the pancreatic duct as indicated. After 20 hours, the rats were killed, pancreata (open bars) and lungs (solid bars) removed, and tissue MPO activities determined. Data shown are means ± SE from 3–6 independent experiments. *P < 0.05 vs. none; **P < 0.05 vs. AdGFP; ##P < 0.05 vs. Adp65 + AdGFP.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Administration of Adp65 induces pancreatic edema and a transient increase in serum amylase levels. Pancreata were analyzed from control animals (none) and those administered either Adp65 or the negative control adenovirus AdGFP. Animals were injected with virus (3 × 107 pfu) and then killed after 16 hours (hatched bars) or 48 hours (open bars); samples of pancreas and serum were collected and (A) pancreatic edema and (B) serum amylase levels were measured. Pancreatic edema was evaluated by measuring the wet-to-dry weight ratio. Assessment of serum amylase levels used the Phadebas amylase test. For all data, values shown are means ± SEM for 3–6 independent experiments. *P < 0.05 vs. control, **P < 0.05 vs. AdGFP.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Administration of Adp65 causes pathologic alterations in pancreatic histology. Sections of pancreas from (A) control animals, (B) animals administered Adp65 (3 × 107 pfu) for 16 hours (C, at higher magnification), or (D) 48 hours, or animals administered AdGFP (3 × 107 pfu) for (E) 16 or (F) 48 hours were prepared for histologic examination (bars = 100 μm). In the higher-magnification micrograph from animals administered Adp65 for 16 hours (C), the open arrows indicate polymorphonuclear leukocytes and closed arrows indicate acinar cells with conspicuous vacuoles. (G) Damage to pancreatic acinar cells was quantitated in histologic sections after 16 hours (hatched bars) or 48 hours (open bars). Five random fields from each of 3 separate animals were evaluated for the percentage of acinar cells with a healthy appearance and those acinar cells bearing signs of damage such as the presence of vacuoles or loss of polarity. Data shown is mean ± SEM for 3 independent experiments. *P < 0.05 vs. control, **P < 0.05 vs. AdGFP.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Pancreatic administration of Adp65 increases neutrophilic infiltration of the lung. Lungs from (A) control animals, (B) those administered AdGFP (3 × 107 pfu), or (C) Adp65 (3 × 107 pfu) were removed after 48 hours and processed for routine histologic analysis (bars = 100 μm). (D) Neutrophilic infiltration was quantitated 48 hours after viral administration by measuring MPO activity. Values shown are means ± SEM for 3–6 independent experiments. *P < 0.05 vs. control, **P< 0.05 vs. AdGFP.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Administration of Adp65 does not specifically activate pancreatic trypsinogen. Pancreata from animals administered Adp65 (3 × 107 pfu) or AdGFP (3 × 107 pfu) for either 16 hours (hatched bars) or 48 hours (open bars) were analyzed for activation of trypsinogen to trypsin by measuring the level of active trypsin. As a positive control, animals were injected intravenously with a bolus of cerulein (10 μg/kg body wt), and this was repeated once 1 hour later; these animals were then killed after 4 hours (CR). Values shown are means ± SEM (n = 6–8). *P < 0.05 vs. control.
Gastroenterology  , DOI: ( /gast )
Copyright © 2002 American Gastroenterological Association Terms and Conditions