title: NecroX-5 suppresses IgE/Ag-stimulated anaphylaxis and mast cell activation by regulating the SHP-1-Syk signaling module.
abstract: BACKGROUND
  IgE/Ag-stimulated mast cells release various pro-allergic inflammatory mediators, including histamine, eicosanoids, and pro-inflammatory cytokines. NecroX-5, a cell permeable necrosis inhibitor, showed cytoprotective effects in both in vitro and in vivo models. However, the anti-allergic effect of NecroX-5 has not yet been investigated. The aims of this study were to evaluate the anti-allergic activity of NecroX-5 in vivo and to investigate the underlying mechanism in vitro.
  
  
  METHODS
  The anti-allergic activity of NecroX-5 was evaluated in vitro using bone marrow-derived mast cells (BMMCs) and IgE receptor-bearing RBL-2H3 or KU812 cells and in vivo using a mouse model of passive anaphylaxis. The levels of histamine, eicosanoids (PGD2 and LTC4 ), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were measured using enzyme immunoassay kits. The mechanism underlying the action of NecroX-5 was investigated using immunoblotting, immunoprecipitation, and gene knockdown techniques.
  
  
  RESULTS
  NecroX-5 markedly inhibited mast cell degranulation and the synthesis of eicosanoids, TNF-α, and IL-6 by suppressing the activation of Syk, LAT, phospholipase Cγ1, MAP kinases, the Akt/NF-κB pathway, and intracellular Ca(2+) mobilization via the activation of phosphatase SHP-1. Oral administration of NecroX-5 effectively suppressed mast cell-dependent passive cutaneous and systemic anaphylactic reactions in a dose-dependent manner.
  
  
  CONCLUSIONS
  NecroX-5 might be a potential candidate for the development of a novel anti-allergic agent that suppresses IgE-dependent mast cells signaling.
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk, Korea.
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk, Korea.
authors.forename: O
authors.initials: O
authors.lastname: Kwon
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk, Korea.
authors.forename: D Y
authors.initials: DY
authors.lastname: Kim
authors.affiliation: Lipid Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Taketomi
authors.affiliation: Lipid Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Murakami
authors.affiliation: College of Pharmacy, Yeungnam University, Gyeongsan, Gyeongbuk, Korea.
authors.forename: H W
authors.initials: HW
authors.lastname: Chang
mesh_terms: D000707:Anaphylaxis / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism*; D000818:Animals; D000941:Antigens / Q000276:immunology*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D015550:Cell Degranulation / Q000187:drug effects / Q000276:immunology; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D004195:Disease Models, Animal; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology*; D007073:Immunoglobulin E / Q000276:immunology*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D017997:Leukotriene C4 / Q000096:biosynthesis; D008297:Male; D008407:Mast Cells / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D015230:Prostaglandin D2 / Q000378:metabolism; D011485:Protein Binding; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D013450:Sulfones / Q000494:pharmacology*; D000072377:Syk Kinase
chemical_list: D000941:Antigens; D016207:Cytokines; D006576:Heterocyclic Compounds, 4 or More Rings; D047908:Intracellular Signaling Peptides and Proteins; C574766:NecroX-5; D013450:Sulfones; D017997:Leukotriene C4; D007073:Immunoglobulin E; D001094:Arachidonate 5-Lipoxygenase; D051546:Cyclooxygenase 2; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; C000606897:Syk protein, mouse; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D015230:Prostaglandin D2; D002118:Calcium
keywords: NecroX-5; Src homology 2 domain-containing phosphatase-1; anti-allergy compound; eicosanoids; spleen tyrosine kinase