Modulatory effects of phytoglycoprotein (75 kDa) on allergic inflammatory cytokines in Di(2-ethylhexyl) phthalate (DEHP)-stimulated RBL-2H3 cells.
This study investigated the inhibitory effect of a glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein) on di(2-ethylhexyl) phthalate (DEHP)-induced mast cell degranulation and related signaling cascade in RBL-2H3 cells. This experiment evaluated the intracellular Ca(2+) level, and the activities of protein kinase C (PKC), mitogen-activated protein kinase (MAPK), transcription factor, and the cytokines in DEHP-treated RBL-2H3 cells. Our results revealed that the CTB glycoprotein in the presence of DEHP inhibits the release of histamine and expression of interleukin (IL)-4, IL-6, and TNF-alpha in RBL-2H3 cells. We also found that the CTB glycoprotein inhibits the intracellular Ca(2+) level, translocation of PKC from cytosol to membrane and the phosphorylation of ERK1/2 in cells. Moreover, the CTB glycoprotein (100 microg/ml) has suppressive effects on transcriptional activation of nuclear factor (NF)-kappaB in DEHP-treated RBL-2H3 cells. The activation of NF-kappaB was collectively blocked by treatment with PKC inhibitor (staurosporine) as well as ERK1/2 inhibitor (PD98059), respectively. The results from these experiments indicated that the CTB glycoprotein inhibits release of histamine and expressions of IL-4, IL-6, and TNF-alpha via down regulations of PKC/MAPK and NF-kappaB on the stage of mast cell degranulation induced by DEHP. Moreover, oral administration of CTB glycoprotein (10-20 mg/kg) inhibited compound 48/80-mediated systemic reaction in mice. In conclusion, we speculated that the CTB glycoprotein might be one component for preparation of health supplements for prevention of allergic immune disorders.
Oh
Phil-Sun
PS
Molecular Biochemistry Laboratory, Biotechnology Research Institute and Center for the Control of Animal Hazards Using Biotechnology (BK21), Chonnam National University, 300 Yongbong-Dong, Gwang-ju 500-757, South Korea.
Lim
Kye-Taek
KT
D000707:Anaphylaxis / Q000139:chemically induced / Q000517:prevention & control; D000818:Animals; D018926:Anti-Allergic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D015550:Cell Degranulation / Q000187:drug effects; D016207:Cytokines / Q000187:drug effects / Q000378:metabolism; D004051:Diethylhexyl Phthalate / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006023:Glycoproteins / Q000494:pharmacology*; D006967:Hypersensitivity / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism*; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism*; D008407:Mast Cells / Q000187:drug effects* / Q000378:metabolism; D029586:Moraceae / Q000737:chemistry* / Q000276:immunology; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D003189:p-Methoxy-N-methylphenethylamine / Q000633:toxicity
D018926:Anti-Allergic Agents; D016207:Cytokines; D006023:Glycoproteins; D003189:p-Methoxy-N-methylphenethylamine; D004051:Diethylhexyl Phthalate; D011493:Protein Kinase C; D048049:Extracellular Signal-Regulated MAP Kinases