Intracellular trypsin induces pancreatic acinar cell death but not NF-κB activation

Baoan D Ji, Sebastian Gaiser, Xueqing Chen, Stephen A. Ernst, Craig D. Logsdon

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Premature intracellular activation of the digestive enzyme trypsinogen is considered to be the initiating event in pancreatitis. However, the direct consequences of intracellular trypsin activity have not previously been examined. In the current study, a mutant trypsinogen (paired basic amino acid cleaving enzyme (PACE)-trypsinogen), which is activated intracellularly by the endogenous protease PACE, was developed. This new construct allowed for the first time direct examination of the effects of intracellular trypsin on pancreatic acinar cells. We found that PACE-trypsinogen was expressed in the secretory pathway and was activated within acinar cells. Expression of PACE-trypsinogen induced apoptosis of HEK293 cells and pancreatic acinar cells, as indicated by histology, DNA laddering, PARP cleavage, and caspase-3 activation. Cell death was blocked by the trypsin inhibitor Pefabloc but not by the pancaspase inhibitor benzyloxycarbonyl-VAD, indicating that caspase-independent pathways were also involved. However, intracellular trypsin had no significant effect on the activity of the proinflammatory transcription factor NF-κB. In contrast, extracellular trypsin caused cell damage and dramatically increased NF-κB activity. These data indicate that localization of active trypsin determines its effects on pancreatic acinar cells. This new model will greatly improve our understanding of the role of active trypsin in pancreatitis and its associated inflammatory response.

Original languageEnglish (US)
Pages (from-to)17488-17498
Number of pages11
JournalJournal of Biological Chemistry
Volume284
Issue number26
DOIs
StatePublished - Jun 26 2009
Externally publishedYes

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Acinar Cells
Trypsinogen
Cell death
Trypsin
Furin
Cell Death
Chemical activation
Pancreatitis
Histology
Enzyme Activation
Trypsin Inhibitors
HEK293 Cells
Secretory Pathway
Caspases
Caspase 3
Peptide Hydrolases
Transcription Factors
Cells
Apoptosis
DNA

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Intracellular trypsin induces pancreatic acinar cell death but not NF-κB activation. / Ji, Baoan D; Gaiser, Sebastian; Chen, Xueqing; Ernst, Stephen A.; Logsdon, Craig D.

In: Journal of Biological Chemistry, Vol. 284, No. 26, 26.06.2009, p. 17488-17498.

Research output: Contribution to journalArticle

Ji, Baoan D ; Gaiser, Sebastian ; Chen, Xueqing ; Ernst, Stephen A. ; Logsdon, Craig D. / Intracellular trypsin induces pancreatic acinar cell death but not NF-κB activation. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 26. pp. 17488-17498.
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