NITRIC OXIDE SYNTHESIS,NITROSOGLUTATHIONE FORMATION AND POSSIBLE GENERATION OF PEROXYNITRITE IN THE CULTURE OF ALVOLAR MACROPHAGES WITH ASBESTOS OR MAN-MADE MINERAL FIBERS.

当用铬石棉或人造矿物纤维（MMMF）培养时，通常用硝酸氧化物（NO）和超氧化物阴离子（O_2-）的显著数量进行培养时，用过氧化物酶（OOO-）的可能形成及其分解为羟基（·OH），其中含有各种细胞和组织的毒性。这些发现强烈地认为，所有NO、O_2-、OOO-和OH衍生出的OOO-分解反映在石棉-或MMMF诱导的肺疾病的发展中。Moreover，这样的发现也是一致的，在先前的研究中，对大鼠体内石棉的无合成和历史病理学研究中发现的，对石棉或MMMF也有明显的反应，特别是在细胞中形成硝酸盐化合物的硝酸盐（GSH），形成硝酸盐化合物的硝酸盐（GS-NO）。如预期的，GS-NO可以在与石棉或MMMFS的AMs的文化中检测到，但在细胞层中检测到的很难。这一发现的索引，即GS-NO，已被释放成类似于氧化物化谷氨酰胺的提取空间。与石棉或MMMFs相比，条件媒体中AMs的文化中GS-NO的数量很小，而且可以与AMs单独控制文化中的AMs相比。However， AMs被培养成细菌内毒素， LPS可显著增强的NO合成，以在条件媒体中释放一个较大的GS-NO数量。因此，矿物纤维被添加到AMs的文化中，没有增加GS-NO的数量，因此，在这种文化中，AMs的GSH合成系统不受矿物纤维的影响，而中性GSH水平并不明显降低。

When cultured with chrysotile asbestos or man-made mineral fibers(MMMF), rat alveolar macrophages (AMs) concomitantly produced in a significant amount of nitric oxide(NO) and superoxide anion (O_2-), indicating a possible formation of peroxynitrite (ONOO-) and its decomposition to hydroxyl radical (・OH), which is very toxic to various cells and tissues. These findings strongly suggest that all of NO, O_2- , ONOO- and OH derived from decomposition of ONOO- are implicated in the development of asbestos- or MMMF-induced lung diseases. Moreover, such findings were also consistent with NO synthesis and histological pathologies observed in the previous studies of rat instilled asbestos into the trachea.NOsythesized by AMs cultured with asbestos or MMMF also readily reacts with thiol compounds, especially with reduced glutathione (GSH) in the cells to form nitrosocompounds such as nitrosoglutathione (GS-NO). As expected, GS-NO was detectable in the conditioned media from the culture of AMs with asbestos or MMMFS, but hardly detected in the cell layer. This finding indicates that GS-NO is released into extracellular space similarly to oxidized glutathione. The amount of GS-NO in the conditioned media from culture of AMs with asbestos or MMMFs was small and comparable to that from the control culture of AMs alone. However, AMs cultured with bacterial endotoxin, LPS remarkably enhanced NO synthesis to release a large amount of GS-NO in the conditioned media. Thus, mineral fibers added into the culture of AMs did not increase the amount of GS-NO.It therefore follows that as far as GSH synthesis systems of AMs are not affected in the culture with mineral fibers, intracellular GSH level is not significantly decreased.