| Co-Investigator(Kenkyū-buntansha) |
MAKIYAMA Kiyoshi Nihon University, School of Medicine, Assistant Professor, 医学部, 講師 (00139172)
INOUE Hajime Nihon University, School of Medicine, Assistant Professor, 医学部, 講師 (60193603)
茂木立 学 日本大学, 医学部, 助手 (50366598)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Research Abstract |
NO-superoxide system is a defensive system to protect against various pathogens by injurious peroxynitrite that is produced in the system when NO reacts with superoxide, and it attacks tyrosine residues resulting in forming nitrotyrosine. Nasal mucosal epithelial cells were separated from mucosal specimen, which was obtained during nasal operations, and cultured with epidermal growth factor for one week or more periods, exchanging culture medium every two days. The cultured human nasal epithelial cells were challenged with cytomix (the mixture of 10ng/mL IL-β, 10ng/mL TNF-α and 10ng/mL IFN-γ), and mRNAs of NO-superoxide system related enzymes were studied by real time-PCR. As the results, the primary cytokines significantly induced iNOS mRNA, xanthine oxidase mRNA and also COX-2 mRNA. The nasal mucosal specimen were immediately fixed in 4% paraform aldehyde in 0.1M PBS, pH 7.5, and thin specimens from the paraffin blocks were studied for immunoreactivities for iNOS, xanthine oxidase, superoxide dismutase, and nitrotyrosine. The epithelial cells and inflamed cells of nasal polyps showed imnunoreactivity for iNOS, nitrotyrosine, superoxide dismutase and xanthine oxidase. Otherwise, of inferior turbinate mucosal gland cells, epithelial cells and inflamed cells showed immunoreactivity for iNOS, nitrotyrosine, superoxide dismutase and xanthine oxidase. A part of nitrotyrosine positive cells were swollen showing morphological change suggesting cell injury, although nitrotyrosine positive cells without morphological alteration suggesting a certain dysfunctions.
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