A STUDY ON MECHANISM OF THE DELAYED CELL IMPAIREMANT OF THE AIRWAY MUCOSA - RELEVANCE OF THE LIPID MEDIATORS TO NITRIC OXIDE AND SUPER OXIDE -

气道粘膜迟发性细胞损伤机制的研究-脂质介质与一氧化氮和超氧化物的相关性-

• 批准号: 12671684
• 负责人: HISAMATSU Ken-ichi
• 金额: $ 2.24万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671684/
• 关键词: human nasal mucosa; delayed mucosal cell impairment; lipid mediator; nitric oxide; superoxide; nitrotyrosine; leukotriene D_4; platelet activating factor; 鼻アレルギー; 気道粘膜細胞障害; ロイコトリエンD4; ECP

The aim of this project is to clarify nasal mucosal impairment in nasal allergy patients and to get clues to the solution of the mechanisms of lipids mediators induced delayed impairment of airway mucosa.I. The following substances were measured in the nasal lavage fluids from nasal allergy patients, and urea was also measured in sera and the lavage fluids for conversion in the nasal secretions. Results : Followings showed significant relationships ; 1) alkariphosphatase activity (ALP) and LDH activity (LDH), 2) ECP and LDH, 3) ECP and ALP, 4) NO and nitrotyrosine (NT), 5) NT and ALP, although followings did not show significant relationships, 6) NT and LDH, 7) ECP and NT. These suggested (1) impairment of allergic nasal mucosa, (2) nasal mucosal cell membrane impairment and flux of LDH from cytoplasm, (3) cell membrane impairment by NO and superoxide system, (4) capable difference between fluxes of ALP from cell membrane and ADH from cytoplasm, (5) NO and superoxide protein impairment system is independent from eosinophil granule protein injurious system.II. LDH, ALP, NO and NT were measured in the medium before and after exposure of 10^<-7> M, 10^<-8> MLTD_4 or PAF to nasal mucosa obtained at a time of surgery. Results : These substances showed tend to increase in progress. (2) Leukotriene receptor antagonist, pranlukast, inhibited LTD_4 induced these increases in the medium. (3) The nasal mucosa exposed to lipid mediators were diffusely stained by anti-NT antibody compared to the control. (4) NT was detected in the culture medium with human bronchial epithelial cells following exposure to LTD_4 or PAF after 24 hours. These data suggest that NT detected in the nasal lavage fluids may be attributable from not only epithelial cells but also other mucosal component cells. For the further investigation of the mechanism of the lipid mediator induced delayed mucosal cell impairment, mRNA level of xanthine oxidase and nitric oxside synthase should be studied.

本课题旨在阐明鼻变态反应患者的鼻黏膜损害，并为解决脂质介质诱导的气道黏膜迟发性损害的机制提供线索。在来自鼻变态反应患者的鼻灌洗液中测量了以下物质，并且还在血清和灌洗液中测量了尿素以用于在鼻分泌物中的转化。结果如下：结果表明：1）碱性磷酸酶活性（ALP）与LDH活性（LDH）、2）ECP与LDH、3）ECP与ALP、4）NO与硝基酪氨酸（NT）、5）NT与ALP、6）NT与LDH、7）ECP与NT均呈显著相关。提示：（1）过敏性鼻粘膜损伤;（2）鼻粘膜细胞膜损伤和胞浆LDH释放;（3）NO和超氧化物系统损伤细胞膜;（4）细胞膜ALP释放和胞浆ADH释放存在差异;（5）NO和超氧化物蛋白损伤系统独立于嗜酸性粒细胞颗粒蛋白损伤系统。分别用10 μ <-7>M、10 μ M <-8>MLTD_4或PAF作用于鼻粘膜，测定其作用前后培养液中LDH、ALP、NO和NT的含量。结果：这些物质在病程中呈增加趋势。(2)白三烯受体拮抗剂pranlukast可抑制LTD_4诱导的上述增加。(3)与对照组相比，暴露于脂质介质的鼻粘膜被抗NT抗体弥漫染色。(4)LTD_4或PAF作用于人支气管上皮细胞24小时后，培养液中可检测到NT。这些数据表明，鼻灌洗液中检测到的NT可能不仅来自上皮细胞，而且来自其他粘膜成分细胞。为了进一步研究脂质介质诱导迟发性粘膜细胞损伤的机制，需要对黄嘌呤氧化酶和一氧化氮合酶的mRNA水平进行研究。

项目成果

Inoue H, Ando K, Wakisaka N, et al.: "Effect of nitric oxide synthase inhibitors on vascular hyperpermeability with thermal injury in mice"NITRIC OXIDE. 5. 334-342 (2001)

Inoue H、Ando K、Wakisaka N 等人：“一氧化氮合酶抑制剂对小鼠热损伤血管通透性过高的影响”一氧化氮。

Inoue H, Hisamatsu K, et al.: "Determination of Nitrotyrosine and Related Compounds in Biological Specimens by Competitive Enzyme Immunoassay"NITRIC OXIDE : Biology and Chemistry.

Inoue H、Hisamatsu K 等人：“通过竞争性酶免疫测定法测定生物样本中的硝基酪氨酸和相关化合物”一氧化氮：生物学和化学。

Inoue H, Hisamatsu K, et al.: "Detection of nitrotyrosine and related compounds in biological specimens by competitive enzyme immunoassay"NITRIC OXIDE : Biology and Chemistry. (In print).

Inoue H、Hisamatsu K 等人：“通过竞争性酶免疫测定法检测生物样本中的硝基酪氨酸和相关化合物”一氧化氮：生物学和化学。

Inoue H, Hisamatsu K, et al.: "Rapid and simple determination of histamine-N-methyl transferase activity by high-performance liquid chromatography with UV determination"Mediators of Inflammation. 10. 273-277 (2001)

Inoue H、Hisamatsu K 等人：“通过高效液相色谱法和紫外测定快速简单地测定组胺-N-甲基转移酶活性”炎症介质。

Hisamatsu K, Nakajima M: "Pranlukast protects leukotriene C_<4-> and D_4-induced epithelial cell impairment of the nasal mucosa in vitro"Life Sciences. 67. 2767-2773 (2000)

Hisamatsu K，Nakajima M：“Pranlukast 保护白三烯 C_4-> 和 D_4 诱导的体外鼻粘膜上皮细胞损伤”生命科学。