Korean J Anesthesiol.  2012 Dec;63(6):540-546. 10.4097/kjae.2012.63.6.540.

Urinary trypsin inhibitor attenuates lipopolysaccharide-induced neutrophil activation

• Lee SH
• Kim HJ
• Han HJ
• Li M
• Kwak SH
• Park S

• KMID: 2236256
• DOI: http://doi.org/10.4097/kjae.2012.63.6.540

Abstract

BACKGROUND
Urinary trypsin inhibitor (UTI), which is speculated to have anti-inflammatory effects, is one of serine protease inhibitors found in human urine and blood. The present study was conducted to clarify the effect of urinary trypsin inhibitor (UTI) on human neutrophil activation and its intracellular signaling mechanism in vitro.
METHODS
To assess the possible interactions between UTI and lipopolysaccharide (LPS) in neutrophil activation, neutrophils from human blood were incubated with varying concentrations of UTI (1, 10, 100, 1,000 and 10,000 U/ml) plus LPS (100 ng/ml) or LPS alone in 24-well plates (5 x 106 cells/well). We measured protein levels for interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-alpha) using enzyme-linked immunosorbent assay (ELISA) kits after 4 hours of incubation period. To elucidate the intracellular signaling pathway, we also measured the levels of phosphorylation of p38, ERK1/2 and JNK via Western blot analysis. Moreover, the nuclear levels of nuclear factor-kappa B (NF-kappaB) were determined with electrophoretic mobility shift assays (EMSA).
RESULTS
UTI decreased the expression of inflammatory cytokines, including TNF-alpha and IL-6, and activation of intracellular signaling pathways, such as JNK, but not P38, ERK1/2 and nuclear translocation of NF-kappaB.
CONCLUSIONS
UTI can attenuate LPS-induced neutrophil responses and may partially contribute to the treatment of neutrophil-mediated inflammatory diseases.

Keyword

Cytokines; Mitogen activated protein kinases; Neutrophils; Urinary trypsin inhibitor

MeSH Terms

Blotting, Western
Cytokines
Electrophoretic Mobility Shift Assay
Enzyme-Linked Immunosorbent Assay
Glycoproteins
Humans
Interleukin-6
Interleukins
Mitogen-Activated Protein Kinases
Neutrophil Activation
Neutrophils
Phosphorylation
Serine Proteinase Inhibitors
Trypsin
Tumor Necrosis Factor-alpha
Cytokines
Glycoproteins
Interleukin-6
Interleukins
Mitogen-Activated Protein Kinases
Serine Proteinase Inhibitors
Trypsin
Tumor Necrosis Factor-alpha