Oxidative Stress of Asbestos or its Substitute to the Lung

石棉或其替代品对肺的氧化应激

基本信息

批准号：
12470103
负责人：
IGUCHI Hiroshi
金额：
$ 6.53万
依托单位：
Hyogo College of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

1. Alveolar macrophages of rats, RAW264.7 or J774 cells which is macrophage cell line established from mouse, induced nitric oxide(NO) Synthase(NOS), and increased the prduction of NO in the culture with asbestos or man-made mineral fiber(MMMF), such as glass fiber, ceramic fiber, rock wool or slug wool by way of substitute for asbestos.2. Increase in NO Synthesis was confirmed in the culture of AM from rats instilled asbestos into trachea.3. Generation of superoxide anion, O_2^- as well as NO was significantly enhanced in the culture of AM, RAW264.7 or J774 with asbestos or MMMF.4. When cultured with asbestos or MMMF, RAW264.7, AM or J774 decreased significantly intracellular glutathione levels in both reduced-and ozxdized-form.5. Taken altogether, these findings imply that oxidative stress in AM, RAW264.7 or J774 was strongly euhanced on exposure to asbestos or MMMF.6. It is well known that NO or peroxynitrite ONOO which is product of NO and O_2^- causes a nitration reaction with free thiol group in protein or wih reduced-form glutathione(GSH). These reaction products are RS-NO or GS-NO. We succeeded in developing a simple and new method for measuremert of RS-NO or GS-NO by using fluorescent dye. Using this method, we found an increased amount of GS-NO or RS-NO in conditioned media of AM, RAW264.7 or J774 cells cultured with asbestos or MMMF. This increase reflects at least in part a decrease in glutathione level in the cultured cells, suggesting the enhancement of oxidative stress or change of protein functions.7. Moreover, when cultured with asbestos or MMMF, AM, RAW264.7 or J774 responded to increase the production of proin- flammatory cytokines such as TNF-α, IL-1β, IL-6. The increases were found in BALF (Bronchoalveolar lavage fluid) from rats inntilled asbestos into trachea. These findings suggeet that collagen synthesis increases and will progress to the development of pneumoconiosis, that is, lung fibrosis soon or later.

1. 大鼠肺泡巨噬细胞，RAW264.7或J774细胞（从小鼠建立的巨噬细胞系），诱导一氧化氮（NO）合酶（NOS），并在石棉或人造矿物纤维培养物中增加NO的产生(MMMF)，如玻璃纤维、陶瓷纤维、岩棉或块状棉替代石棉。2．在培养物中证实了NO合成的增加。气管内注入石棉的大鼠AM。3.AM、RAW264.7或J774与石棉或MMMF培养物中超氧阴离子、O_2^-和NO的产生显着增强。4． RAW264.7、AM 或 J774 均显着降低还原形式和 ozxdized 形式的细胞内谷胱甘肽水平。5。总体而言，这些发现表明，接触石棉或 MMMF 后，AM、RAW264.7 或 J774 中的氧化应激会大大增强。 6 众所周知，NO 或过氧亚硝酸盐 ONOO（NO 和 O_2^- 的产物）会引起硝化反应。与蛋白质中的游离硫醇基团或与还原型谷胱甘肽（GSH）反应的产物是RS-NO或GS-NO。使用荧光染料测量 RS-NO 或 GS-NO 的简单而新的方法使用该方法，我们发现用 AM、RAW264.7 或 J774 细胞培养的条件培养基中 GS-NO 或 RS-NO 的量增加。这种增加至少部分反映了培养细胞中谷胱甘肽水平的降低，表明氧化应激的增强或蛋白质功能的变化。7。石棉或 MMMF、AM、RAW264.7 或 J774 会增加促炎性细胞因子（如 TNF-α、IL-1β、IL-6）的产生，这些增加在注入石棉的大鼠 BALF（支气管肺泡灌洗液）中发现。这些发现表明胶原蛋白合成增加并将发展为尘肺病，即肺。纤维化迟早会发生。