Shi Tingming, Duffin Rodger, Borm Paul J A, Li Hui, Weishaupt Christel, Schins Roel P F
Institut für Umweltmedizinische Forschung an der Heinrich-Heine University Düsseldorf gGmbH, auf'm Hennekamp 50, D-40225 Düsseldorf, Germany.
Environ Res. 2006 May;101(1):18-24. doi: 10.1016/j.envres.2005.09.005. Epub 2005 Nov 17.
Exposure to ambient particulate matter (PM) has been reported to be associated with increased respiratory, cardiovascular, and malignant lung disease. Previously we have shown that PM can induce oxidative DNA damage in A549 human lung epithelial cells. The aims of the present study were to investigate the variability of the DNA-damaging properties of PM sampled at different locations and times and to relate the observed effects to the hydroxyl-radical (OH)-generating activities of these samples. Weekly samples of coarse (10-2.5 microm) and fine (<2.5 microm) PM from four sites (Nordrheim Westfalen, Germany) were analyzed for hydrogen-peroxide-dependent OH formation using electron paramagnetic resonance and formation of 8-hydroxydeoxyguanosine (8-OHdG) in calf thymus DNA using an immuno-dot-blot assay. DNA strand breakage by fine PM in A549 human lung epithelial cells was quantified using the alkaline comet assay. Both PM size distribution fractions elicited OH generation and 8-OHdG formations in calf thymus DNA. Significantly higher OH generation was observed for PM sampled at urban/industrial locations and for coarse PM. Samples of fine PM also caused DNA strand breakage in A549 cells and this damage could be prevented using the hydroxyl-radical scavengers 5,5-dimethyl-1-pyrroline-N-oxide and dimethyl sulfoxide. The observed DNA strand breakage appeared to correlate with the hydroxyl-radical-generating capacities of the PM samples but with different profiles for rural versus urban/industrial samples. In conclusion, when considered at equal mass, OH formation of PM shows considerable variability with regard to the sampling location and time and is correlated with its ability to cause DNA damage.
据报道,暴露于环境颗粒物(PM)与呼吸系统、心血管系统疾病及恶性肺部疾病的增加有关。此前我们已经表明,PM可诱导A549人肺上皮细胞中的氧化性DNA损伤。本研究的目的是调查在不同地点和时间采集的PM的DNA损伤特性的变异性,并将观察到的效应与这些样品的羟基自由基(OH)生成活性联系起来。使用电子顺磁共振分析了来自四个地点(德国北莱茵-威斯特法伦州)的粗颗粒物(10 - 2.5微米)和细颗粒物(<2.5微米)的每周样品中过氧化氢依赖性OH的形成,并使用免疫斑点印迹法分析了小牛胸腺DNA中8-羟基脱氧鸟苷(8-OHdG)的形成。使用碱性彗星试验对A549人肺上皮细胞中细PM引起的DNA链断裂进行了定量。两种PM粒径分布级分均能在小牛胸腺DNA中引发OH生成和8-OHdG形成。在城市/工业地点采集的PM以及粗PM中观察到显著更高的OH生成。细PM样品也会导致A549细胞中的DNA链断裂,并且这种损伤可以使用羟基自由基清除剂5,5-二甲基-1-吡咯啉-N-氧化物和二甲基亚砜来预防。观察到的DNA链断裂似乎与PM样品的羟基自由基生成能力相关,但农村样品与城市/工业样品的情况不同。总之,当以等量质量考虑时,PM的OH形成在采样地点和时间方面表现出相当大的变异性,并且与其引起DNA损伤的能力相关。