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，RAW264.7或J774细胞的肺泡巨噬细胞，该细胞是由小鼠建立的，诱导的一氧化氮（NO）合酶（NOS），并增加了与石棉或人为矿物纤维（MMMF）的培养物中NO培养物的production，例如玻璃纤维，例如玻璃纤维，摩克式羊毛材料，sl sl.sl.sl. sloce fiber，slob fiber，sl sl sl oct sloct sl occeb ock slo slo sl.在AM的培养物中证实了无合成的增加，将石棉灌输到气管中。3。在AM，RAW264.7或J774的培养物中产生的超氧化阴离子O_2^ - 与石棉或MMMF.4的培养物显着增强。当用石棉或MMMF培养时，RAW264.7，AM或J774在降低的ozxdized形式下，AM或J774培养可显着的细胞内谷胱甘肽水平。5。这些发现完全是，AM，RAW264.7或J774中的氧化应激强烈地暴露于石棉或MMMF.6上。众所周知，NO和O_2^的乘积NO或过氧亚硝酸盐 - 在蛋白质或WIH降低形式的谷胱甘肽（GSH）中引起硝化反应。这些反应产物是RS-NO或GS-NO。我们通过使用荧光染料成功开发了一种简单的新方法来测量RS-NO或GS-NO。使用这种方法，我们在AM，RAW264.7或用石棉或MMMF培养的条件介质中发现了增加的GS-NO或RS-NO。这种增加至少反映了培养细胞中谷胱甘肽水平的部分降低，这表明氧化应激的增强或蛋白质功能的变化7。此外，当用石棉或MMMF培养时，AM，RAW264.7或J774响应以增加促炎性细胞因子的产生，例如TNF-α，IL-1β，IL-6。从大鼠含石棉进入气管的BALF（支气管肺泡灌洗液）中发现了增加。这些发现表明，胶原蛋白合成会增加，并将发展为肺炎的发展，即很快或以后的肺纤维化。