NITRIC OXIDE RESPONSES OF MAST CELLS AND EOSINOPHILS

在哺乳动物组织中由L-精氨酸产生一氧化氮（NO）是一种生物学过程。 成为一个紧张的调查领域。 不同的生理作用是 因此被证明或假设，表明NO是重要的 血管反应中效应分子，血小板活性， 神经传递和炎症。 最明确的角色是 来自炎性/免疫活性细胞的NO是细胞毒性的。 活化巨噬细胞的作用。 本建议的核心是 几乎未研究的NO从肥大细胞和嗜酸性粒细胞产生， 与过敏性疾病、关节炎、神经源性 炎症和哮喘的潜在肺部炎症。 在 与NO的稳态作用相反，这些物质过度释放NO， 活化的炎性细胞可能有助于病理生理学 某些炎症/过敏性疾病的症状。 使用建立在 体外肥大细胞和嗜酸性粒细胞培养模型，l-精氨酸/NO 将在激活的细胞中探索这一途径。 拟议的具体目标重点 在实验设计，以研究产生NO以下细胞 通过各种信号转导途径刺激。 肥大细胞将 用IgE-抗二硝基酚BSA刺激，单独用佛波酯肉豆蔻酸酯 离子载体A23187、P物质、γ-干扰素、肿瘤坏死因子 α因子和脂多糖。 嗜酸性粒细胞将被刺激， γ-干扰素，肿瘤坏死因子-α，脂多糖， 细菌细胞壁肽，血小板活化因子，佛波醇 肉豆蔻酸酯乙酸酯和P物质。因此，细胞将通过 免疫途径、肽受体刺激和蛋白激酶C activation. 超氧化物及超氧化物歧化酶对NO的调节作用 释放将被探索。 NO亚型选择性药理学抑制剂 NO合酶将用于描述NO合酶同种型的类型 存在于肥大细胞和嗜酸性粒细胞中。 成功完成这些 调查可能会产生有益的结果，在未来的发展新的 治疗炎症性疾病的治疗方法， 危及血管内平衡 NO的升压机制

Generation of nitric oxide (NO) from L-arginine in mammalian tissues is becoming an area of intense investigation. Diverse physiologic roles are thus being demonstrated or postulated suggesting that NO is an important effector molecule in vascular responses, platelet activity, neurotransmission and inflammation. The most clearly established role for NO from inflammatory/immunologically competent cells is the cytotoxic effect of activated macrophages. Central to the present proposal is the virtually unstudied generation of NO from mast cells and eosinophils, both inflammatory cells implicated in allergic disorders, arthritis, neurogenic inflammation and the underlying pulmonary inflammation of asthma. In contrast to the homeostatic roles of NO, its exaggerated release by these activated inflammatory cells may contribute to tahe pathophysiologic symptoms of certain inflammatory/allergic diseases. Using established in vitro mast cell and eosinophil cell culture models, the l-arginine/NO pathway will be explored in activated cells. Proposed specific aims focus on experiments designed to investigate generation of NO following cell stimulation via various signal transduction pathways. Mast cells will be stimulated with IgE-anti-dinitrophenol BSA, phorbol myristate acetate alone and with ionophore A23187, Substance P, gamma-interferon, tumor necrosis factor-alpha and lipopolysaccharide. Eosinophils will be stimulated with gamma-interferon, tumor necrosis factor-alpha, lipopolysaccharide, the bacterial cell wall peptide fMLP, platelet activating factor, phorbol myristate acetate and Substance P. Thus, cells will be stimulated via immunologic pathways, peptide receptor stimulation and protein kinase C activation. Modulatory roles of superoxide and superoxide dismutase on No release will be explored. Isoform-selective pharmacologic inhibitors of NO synthase will be used to describe the type(s) of NO synthase isoforms present in mast cells and eosinophils. Successful completion of these investigations may yield results useful in the future development of new therapeutic approaches to the treatment of inflammatory diseases without compromising homeostatic vascular pressor mechanisms of NO.