Biochemical and Biophysical Research Communications
Proinflammatory effects of pancreatic elastase are mediated through TLR4 and NF-κB☆
Section snippets
Cells and materials
Human myeloid cell lines U-937 and THP-1 were obtained from ECACC (European Collection of Cell Cultures, CAMR, Salisbury, UK). The cells were cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 100 mg/ml streptomycin and 100 U/ml penicillin (Gibco Europe, Paisley, UK). Porcine pancreatic elastase (#324723) and human recombinant tumor necrosis factor-α (TNF-α) (#654205) were purchased from Calbiochem (EMD Biosciences, Darmstadt, Germany). Lipopolysaccharide (LPS, Escherichia
Pancreatic elastase activates several transcription factors
Pancreatic elastase induced activation of NF-κB, AP-1, and NFAT transcription factors tested at 3 h (Fig. 1A). Elastase-induced NF-κB activation was not as rapid as the activation induced by TNF-α or LPS (30 min) but was clearly present at 3 h (Fig. 1B). Pancreatic elastase was non-toxic at the concentration used (1 U/ml). The viability and cell proliferation index (XTT-test) of elastase-stimulated U-937 and THP-1 cells did not differ from controls at 48 h (data not shown).
Pancreatic elastase-induced TNF-α secretion is NF-κB dependent
Elastase-induced NF-κB
Discussion
Pancreatic elastase has been implicated in the activation of inflammatory cells during acute pancreatitis. On one hand, prematurely activated pancreatic elastase has been found in pancreatic tissue in experimental acute pancreatitis [10]. On the other hand, pancreatic elastase has been shown to activate inflammatory cells in vivo and in vitro [4], [5]. Further, pancreatic elastase can be used to induce a human SIRS-like inflammatory condition with associated mortality in experimental animals [6]
Acknowledgments
These studies were supported by Sigrid Juselius Foundation and Helsinki University Central Hospital Research Grant (EVO).
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Abbreviations: AP-1, activating protein-1; EMSA, electrophoretic mobility shift assay; IκB, inhibitory-κB; NFAT, nuclear factor of activated T-cells; NF-κB, nuclear factor-κB; PGA1, prostaglandin A1; TLR, Toll-like receptor; TNF-α, tumor necrosis factor-α.