Huang Yuh-Chin T, Li Zhuowei, Carter Jacqueline D, Soukup Joleen M, Schwartz David A, Yang Ivana V
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27705, USA.
Am J Respir Cell Mol Biol. 2009 Nov;41(5):544-52. doi: 10.1165/rcmb.2008-0064OC. Epub 2009 Feb 27.
Exposure to pollutant particles increased respiratory morbidity and mortality. The alveolar macrophages (AMs) are one cell type in the lung directly exposed to particles. Upon contact with particles, AMs are activated and produce reactive oxygen species, but the scope of this oxidative stress response remains poorly defined. In this study, we determined the gene expression profile in human AMs exposed to particles, and sought to characterize the global response of pro- and antioxidant genes. We exposed AMs obtained by bronchoscopy from normal individuals to Chapel Hill particulate matter of 2.5-microm diameter or smaller (PM(2.5); 1 microg/ml) or vehicle for 4 hours (n = 6 independent samples). mRNAs were extracted, amplified, and hybridized to Agilent human 1A microarray. Significant genes were identified by significance analysis of microarrays (false discovery rate, 10%; P < or = 0.05) and mapped with Gene Ontology in the Database for Annotation, Visualization, and Integrated Discovery. We found 34 and 41 up- and down-regulated genes, respectively; 22 genes (approximately 30%) were involved in metal binding, and 11 were linked to oxidative stress, including up-regulation of five metallothionein (MT)-1 isoforms. Exogenous MT1 attenuated PM(2.5)-induced H2O2 release. PM(2.5) premixed with MT1 stimulated less H2O2 release. Knockdown of MT1F gene increased PM(2.5)-induced H2O2 release. PM(2.5) at 1 microg/ml did not increase H2O2 release. Mount St. Helens PM(2.5) and acid-extracted Chapel Hill PM(2.5), both poor in metals, did not induce MT1F or H2O2 release. Our results show that PM(2.5) induced a gene expression profile prevalent with genes related to metal binding and oxidative stress in human AMs, independent of oxidative stress. Metals associated with PM may play an important role in particle-induced gene changes.
接触污染物颗粒会增加呼吸道疾病的发病率和死亡率。肺泡巨噬细胞(AMs)是肺部直接接触颗粒的一种细胞类型。与颗粒接触后,AMs被激活并产生活性氧,但这种氧化应激反应的范围仍不清楚。在本研究中,我们确定了暴露于颗粒的人AMs中的基因表达谱,并试图描述促氧化和抗氧化基因的整体反应。我们将通过支气管镜从正常个体获得的AMs暴露于直径为2.5微米或更小的教堂山颗粒物(PM(2.5);1微克/毫升)或赋形剂中4小时(n = 6个独立样本)。提取mRNA,进行扩增,并与安捷伦人类1A微阵列杂交。通过微阵列显著性分析(错误发现率,10%;P≤0.05)鉴定显著基因,并在注释、可视化和综合发现数据库中用基因本体进行映射。我们分别发现了34个上调基因和41个下调基因;22个基因(约30%)参与金属结合,11个基因与氧化应激相关,包括5种金属硫蛋白(MT)-1亚型的上调。外源性MT1减弱了PM(2.5)诱导的H2O2释放。与MT-1预混合的PM(2.5)刺激产生的H2O2释放较少。MT1F基因敲低增加了PM(2.5)诱导的H2O2释放。1微克/毫升的PM(2.5)不会增加H2O2释放。圣海伦斯火山的PM(2.5)和酸提取的教堂山PM(2.5),两者金属含量都很低,不会诱导MT1F或H2O2释放。我们的结果表明,PM(2.5)在人AMs中诱导了一个以与金属结合和氧化应激相关的基因占主导的基因表达谱,与氧化应激无关。与PM相关的金属可能在颗粒诱导的基因变化中起重要作用。