Using yeast RNA as a probe for generation of hydroxyl radicals by earth materials
Using yeast RNA as a probe for generation of hydroxyl radicals by earth materials
复制标题
DOI：
10.1021/es052301k
复制
发表时间：
2006-04-15
影响因子：
11.4
通讯作者：
Schoonen, MAA
中科院分区：
文献类型：
--
作者：
Cohn, CA;Laffers, R;Schoonen, MAA
吸入某些类型的颗粒物可导致肺部疾病。这些颗粒的反应性以及部分导致的病理反应由其物理化学性质决定。颗粒诱导活性氧（ROS），特别是体内羟基自由基产生的能力是与肺部疾病发展相关的一种性质。一些矿物质，如石英和石棉，已知会产生羟基自由基并导致肺部疾病，但许多其他矿物质从未进行过测试。在这里，我们描述了一种技术，采用酵母RNA作为探针筛选矿物质产生的羟基自由基。RNA在过氧化氢、亚铁、羟基自由基和几种常见矿物（石英、钠长石、镁橄榄石、铁橄榄石、赤铁矿、磁铁矿、煤和黄铁矿）存在下的稳定性进行了研究。使用3 '-（对氨基苯基）荧光素（APF）来验证ROS的矿物质产生。RNA在过氧化氢、石英和钠长石的存在下是稳定的;而在亚铁、羟基自由基和其他矿物质的存在下则会降解。煤和黄铁矿在RNA降解和羟基自由基产生方面都是最具活性的。这种非细胞技术提供了一种直接的方法来同时比较许多不同的粒子。使用这种方法显示出对RNA的反应性的那些颗粒是进一步体外和可能的体内测试的高优先级候选物。
Inhalation of certain types of particulate matter can lead to lung disease. The reactivity of these particles and, in part, the pathologic responses that result are dictated by their physicochemical properties. The ability of particles to induce the generation of reactive oxygen species (ROS), especially hydroxyl radicals in vivo, is one property that has been correlated to the development of lung disease. Several minerals, such as quartz and asbestos, are known to generate hydroxyl radicals and cause lung disease, but many other minerals have never been tested. Here, we describe a technique employing yeast RNA as a probe to screen for mineral-generated hydroxyl radicals. The stability of RNA in the presence of hydrogen peroxide, ferrous iron, hydroxyl radicals, and several common minerals (quartz, albite, forsterite, fayalite, hematite, magnetite, coal, and pyrite) was examined. 3'-(p-Aminophenyl) fluorescein (APF) was used to verify mineral generation of ROS. RNA is stable in the presence of hydrogen peroxide, quartz, and albite; while it degrades in the presence of ferrous iron, hydroxyl radicals, and the other minerals. Coal and pyrite are the most reactive both in RNA degradation and hydroxyl radical generation. This noncellular technique provides a straightforward way to compare many different particles simultaneously. Those particles showing reactivity toward RNA using this method are high-priority candidates for further in vitro and possibly in vivo tests.