SUPEROXIDE AND BONE RESORPTION

这项提议的广泛的长期目标是理解 破骨细胞介导的骨吸收中的超氧化物。我们的关系 在褶皱的边界空间内产生破骨细胞超氧化物之间 和骨吸收；2.确定超氧化物的作用(S) 在骨骼中产生的是由于在破骨细胞中产生的超氧化物 细胞膜或细胞外或两者兼而有之；3.为了确定细胞因子是否， 干扰素-伽玛，已知可刺激白细胞产生超氧化物 从嗜铬肉芽肿病患者身上，增强破骨细胞- 介导性骨吸收和骨质疏松症小鼠突变体，如果是， 将行动地点本地化。在这些实验中， 在分离的破骨细胞培养中将监测超氧阴离子清除剂 还有头盖骨。比较被排除在细胞外的清道夫 从膜的大小到渗透的低分子质量清除剂 膜，超氧化物对骨吸收的影响部位将是 被追寻。通过使用不同年龄的新生动物，我们将评估 破骨细胞形成率在破骨细胞形成机制中的重要性 氧自由基影响骨吸收。NBT染色将用于 测量和定位个体体内氧自由基产生的变化 破骨细胞。干扰素伽玛将被用来刺激超氧化物 从破骨细胞中分离出破骨细胞。如果干扰素伽玛 刺激破骨细胞介导的骨吸收 骨性动物，超氧化物清除剂将添加到相同的 培养以确定超氧化物的产生对环境的贡献 骨吸收能力增强。这些实验已经 为干扰素伽玛治疗的I/II期试验提供基础 骨腐症患者。

The broad long-tern objective of this proposal is to understand the role of superoxide in osteoclast-mediated bone resorption. Our relationship between osteoclastic superoxide generation within the ruffled border space and bone resorption; 2. to determine if the effect(s) of superoxide produced in bone is due to superoxide generated within the osteoclastic cellular membrane or outside or both; 3. to determine if a cytokine, Interferon Gamma, known too stimulate superoxide generation in leukocytes from patients with chromic granulomatous disease, enhances osteoclast- mediated bone resorption in and osteopetrotic mouse mutant and if so, to localize the site of action. In these experiments, the effects of superoxide scavengers will be monitored in cultures of isolated osteoclasts and calvaria. Comparing a scavenger which is excluded from cellular membranes by size to a low molecular weight scavenger which permeates membranes, the site of superoxide's effects upon bone resorption will be sought. By using neonatal animals of different ages, we shall assess the importance of the rate of osteoclasts formation upon the mechanism by which oxygen radicals affect bone resorption. NBT staining will be used to measure and localize changes in oxygen radical production in individual osteoclasts. Interferon Gamma will be used to stimulate superoxide generation in osteoclasts from osteopetrotic misc. If Interferon Gamma stimulates osteoclast-mediated bone resorption in tissues cultured from osteopetrotic animals, superoxide scavengers will be added to identical cultures to determine the contribution of superoxide generation to the increased bone resorptive capacity. These experiments have already provides the basis for a phase I/II trial of Interferon Gamma to treat osteopetrotic patients.