Pancreas-specific RelA/p65 truncation increases susceptibility of acini to inflammation-associated cell death following cerulein pancreatitis
Hana Algül, Matthias Treiber, Marina Lesina, Hassan Nakhai, Dieter Saur, Fabian Geisler, Alexander Pfeifer, Stephan Paxian, Roland M. Schmid
Hana Algül, Matthias Treiber, Marina Lesina, Hassan Nakhai, Dieter Saur, Fabian Geisler, Alexander Pfeifer, Stephan Paxian, Roland M. Schmid
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Research Article

Pancreas-specific RelA/p65 truncation increases susceptibility of acini to inflammation-associated cell death following cerulein pancreatitis

Abstract

Activation of the transcription factor NF-κB/Rel has been shown to be involved in inflammatory disease. Here we studied the role of RelA/p65, the main transactivating subunit, during acute pancreatitis using a Cre-loxP strategy. Selective truncation of the rela gene in pancreatic exocrine cells led to both severe injury of the acinar cells and systemic complications including lung and liver damage. Our data demonstrated that expression and induction of the protective pancreas-specific acute phase protein pancreatitis-associated protein 1 (PAP1) depended on RelA/p65. Lentiviral gene transfer of PAP1 cDNA reduced the extent of necrosis and infiltration in the pancreata of mice with selective truncation of RelA/p65. These results provide in vivo evidence for RelA/p65 protection of acinar cell death via upregulation of PAP1. Moreover, our data underscore the pancreas-specific role of NF-κB/Rel and suggest multidimensional roles of NF-κB/Rel in different cells and contexts during inflammation.

Authors

Hana Algül, Matthias Treiber, Marina Lesina, Hassan Nakhai, Dieter Saur, Fabian Geisler, Alexander Pfeifer, Stephan Paxian, Roland M. Schmid

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Figure 6

Impaired upregulation of murine PAP1 during AP in relaΔ/Δ mice.

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Impaired upregulation of murine PAP1 during AP in relaΔ/Δ mice. ...
(A and B) Pancreata from relaflox/flox and relaΔ/Δ mice were removed at the indicated times. (A) Total pancreatic RNA (8 μg; n = 2) was labeled and hybridized to Affymetrix MOE430A GeneChips, and pancreas-specific genes were clustered hierarchically. (B) Relative levels of PAP1 mRNA were determined by real-time PCR and expressed as mean ± SD (n = 5). (C) Pancreatic tissues were removed at the indicated times. Whole-tissue extracts were prepared and subjected to Western blot analysis using a newly generated antibody to murine PAP1. (D) Chromatin immunoprecipitation experiments were performed with relaΔ/Δ and relaflox/flox pancreatic tissue at the indicated times after stimulation with cerulein using an antibody to p65. Precipitated DNA was analyzed by PCR using primers surrounding the positions of both κB sites in the respective promoters. PCR was also performed with 2.5% of input chromatin to ensure equal loading. (E) Sections of snap-frozen pancreata and duodenum were prepared and analyzed for PAP1 expression in relaflox/flox and relaΔ/Δ mice in unstimulated pancreas and 12 and 24 hours after the first cerulein injection, respectively. Snap-frozen duodenum served as a positive control, because Paneth cells are known to express PAP1. Signals in the pancreas were localized to the apical regions of acini (arrows), typical for secretory proteins like PAP1.