Inhibitory effect of ZPDC glycoprotein on the expression of inflammation-related cytokines through p38 MAP kinase and JNK in lipopolysaccharide-stimulated RAW 264.7 cells.
OBJECTIVE
This study was carried out to investigate the anti-inflammatory potentials of 24 kDa glycoprotein isolated from Zanthoxylum piperitum DC fruit (ZPDC glycoprotein) in lipopolysaccharide (LPS)-stimulated murine macrophage cell line (RAW 264.7 cells).


METHODS
RAW 264.7 cells were treated with ZPDC glycoprotein (50-200 microg/ml) in presence of LPS (2 microg/ml). The changes of the levels of inflammation-related factors were determined by using Western blot, EMSA, and RT-PCR.


RESULTS
ZPDC glycoprotein has inhibitory effects on the phosphorylation of p38 mitogen-activated protein (MAP) kinase and c-Jun N-terminal kinase (JNK), on the DNA binding activity of activator protein-1 (AP-1), and on the expression of c-Jun and c-Fos in LPS-stimulated RAW 264.7 cells. Interestingly, the DNA binding activity of AP-1 was attenuated by treatment with inhibitors of p38 MAP kinase and JNK. In addition, ZPDC glycoprotein (200 microg/ml) not only diminished the production of superoxide anion, hydrogen peroxide, and nitric oxide, but also suppressed the expression of pro-inflammatory cytokines (IL-1 beta, IL-6, and TNF-alpha) and proteins (iNOS, COX-2, and MMP-9) in LPS- stimulated RAW 264.7 cells.


CONCLUSIONS
The present study demonstrates that ZPDC glycoprotein is a natural anti-oxidant and one of the modulators of pro-inflammatory signal transduction pathways in RAW 264.7 cells.
Lee
S-J
SJ
Molecular Biochemistry Laboratory, Biotechnology Research Institute and Center for the Control of Animal Hazards Using Biotechnology (BK 21), Chonnam National University, 300 Yongbong-Dong, Gwangju, 500-757, South Korea.
Lim
K-T
KT
D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D002460:Cell Line; D016207:Cytokines / Q000276:immunology*; D004791:Enzyme Inhibitors / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D007249:Inflammation / Q000276:immunology*; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000276:immunology / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000276:immunology*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D016877:Oxidants / Q000378:metabolism; D015290:Second Messenger Systems / Q000502:physiology; D032106:Zanthoxylum / Q000737:chemistry; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000975:Antioxidants; D016207:Cytokines; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D008070:Lipopolysaccharides; D016877:Oxidants; D009569:Nitric Oxide; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases