Back to Search
Start Over
Pyrrolidine dithiocarbamate-induced macrophage inflammatory protein-2 gene expression is NF-kappaB-independent but c-Jun-dependent in macrophage cell line RAW 264.7.
- Source :
-
Molecular immunology [Mol Immunol] 2005 Jun; Vol. 42 (10), pp. 1165-75. Date of Electronic Publication: 2005 Jan 07. - Publication Year :
- 2005
-
Abstract
- Pyrrolidine dithiocarbamate (PDTC) is a stable compound that acts as antioxidant or prooxidant, and is widely used to inhibit the activation of NF-kappaB. PDTC was also reported to activate NF-kappaB depending on its dose and metal ions in PC12 cells. In this work, we demonstrated a working mechanism of PDTC and its effects on the proinflammatory cytokine gene expression in a mouse macrophage cell line, RAW 264.7. PDTC alone induced NF-kappaB-independent MIP-2 promoter activation that can be assessed by transient transfection and confocal image analysis. The involvement of AP-1 transcription factor was noticed by promoter deletion/site-specific mutation analysis and electrophoretic mobility shift assay (EMSA). Among three different mitogen-activated protein kinase (MAPK) pathways tested, only the stress-activated protein kinase (SAPK)/Jun N-terminal kinase (JNK) pathway was significantly activated in RAW 264.7 cells after the stimulation with PDTC. Using pathway-specific inhibitors, we found that the SAPK/JNK pathway is clearly associated with PDTC-induced MIP-2 gene expression. Our experimental results indicate that PDTC-induced proinflammatory cytokine expressions are mediated by SAPK/JNK pathway, which activates AP-1.
- Subjects :
- Animals
Blotting, Western
Cell Line
Chemokine CXCL2
DNA Mutational Analysis
Electrophoretic Mobility Shift Assay
Enzyme Activation drug effects
Fluorescein-5-isothiocyanate
Fluorescent Antibody Technique, Indirect
Fluorescent Dyes
Genes, Reporter
Luciferases metabolism
Mice
Microscopy, Confocal
Monokines genetics
Mutagenesis, Site-Directed
Promoter Regions, Genetic
Reverse Transcriptase Polymerase Chain Reaction
Transcription Factor AP-1 metabolism
Antioxidants pharmacology
Gene Expression drug effects
Macrophages drug effects
Monokines metabolism
NF-kappa B antagonists & inhibitors
Proto-Oncogene Proteins c-jun metabolism
Pyrrolidines pharmacology
Thiocarbamates pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 0161-5890
- Volume :
- 42
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Molecular immunology
- Publication Type :
- Academic Journal
- Accession number :
- 15829306
- Full Text :
- https://doi.org/10.1016/j.molimm.2004.11.016