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Peroxynitrite modulates release of inflammatory mediators from guinea pig lung mast cells activated by antigen-antibody reaction

Authors
 Ji Young Kim  ;  Kwang Hoon Lee  ;  Bong Ki Lee  ;  Jai Youl Ro 
Citation
 INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY, Vol.137(2) : 104-114, 2005 
Journal Title
 INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 
ISSN
 1018-2438 
Issue Date
2005
MeSH
Animals ; Antigen-Antibody Reactions ; Antioxidants/pharmacology ; Calcium/metabolism ; Female ; Guinea Pigs ; Histamine Release* ; Leukotrienes/metabolism* ; Lung/cytology ; Lung/immunology* ; Mast Cells/immunology* ; Nitric Oxide/metabolism ; Peroxynitrous Acid/physiology* ; Phospholipases A/metabolism ; Reactive Oxygen Species/metabolism ; Tyrosine/analysis
Keywords
Mast cells ; Peroxynitrite ; Reactive oxygen species ; Antioxidants ; Intracellular Ca2+ level ; Phospholipase A2
Abstract
BACKGROUND: Peroxynitrite (ONOO-), the product of the reaction between the superoxide anion (*O2-) and nitric oxide (NO), is produced during inflammatory disease and may be a major cytotoxic agent. No reports are available as to whether ONOO- generates or modulates inflammatory mediator release from activated guinea pig lung mast cells. In this study, we explored the modulatory role of intracellular ONOO- on inflammatory mediator release (histamine and leukotrienes) from activated mast cells. METHODS: Guinea pig lung mast cells were purified by the enzyme digestion, and by using the rough and discontinuous Percoll density gradients. Mast cells were sensitized with IgG1 (anti-ovalbumin) antibody and challenged with ovalbumin (OVA). The intracellular ROS formation was determined by following the oxidative production of 2', 7'-dichlorofluorescein diacetate (DCFH-DA), dihydrorhodamine 123 (DHR), and anti-nitrotyrosine antibody immunofluorescence. Histamine was assayed using a fluorometric analyzer, leukotrienes by radioimmunoassay, intracellular Ca2+ levels by confocal scanning microscopy, and PLA(2) activity using prelabeling of [3H]arachidonic acid. RESULTS: ROS detected by DCFH-DA weakly increased in mast cells activated with OVA (1.0 g/ml), and the ROS so generated was inhibited by ebselen (50 microM). However, the ROS detected by DHR increased 3-fold under the same conditions. Peroxynitrite scavengers sL-MT, DMTU, and inhibitor FeTPPS inhibited ROS formation but the NADPH oxidase inhibitor diphenyleneiodonium (DPI) only partially inhibited this formation. Dimethyl thiourea (DMTU) and seleno-L-methionine (sL-MT) inhibited the tyrosine nitration of cytosolic proteins, the release of histamine and leukotrienes, Ca2+ influx, and the PLA(2) activity evoked by mast cell activation. CONCLUSION: The data obtained suggests that the ROS generated by the antigen/antibody reaction activated mast cells is ONOO-, and that this modulates the release of inflammatory mediators via Ca2+ -dependent PLA(2) activity.
Full Text
http://www.karger.com/Article/FullText/85465
DOI
10.1159/000085465
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Dermatology (피부과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
Yonsei Authors
Ro, Jai Youl(노재열)
Lee, Kwang Hoon(이광훈)
Lee, Bong Ki(이봉기)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/150661
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