臭氧诱导豚鼠气管支气管上皮细胞DNA链断裂。

S F Ferng, C E Castro, A A Afifi, E Bermúdez, M G Mustafa
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引用次数: 28

摘要

臭氧(O3)是光化学烟雾的主要氧化剂,被认为具有遗传毒性,是一种潜在的呼吸道致癌物或致癌过程的促进剂。由于与上呼吸道粘液的氧化反应,O3反应产物能够渗透到气管支气管上皮(TE)细胞中。O3对TE细胞的致癌作用尤其令人感兴趣,因为大多数先前的研究只关注气管的形态或通透性变化。因此,本研究的目的是以DNA链断裂为指标,研究O3在体内暴露后对TE细胞的潜在遗传毒性。两个月大的雄性Dunkin-Hartley豚鼠,无特定病原体,每组4只,暴露于1.0 ppm O3 0、12、24、48、72或96小时。暴露于过滤空气而不暴露于O3的动物作为对照。在O3暴露后,从每只动物身上取下带有两条主支气管的气管,分离TE细胞,通过DNA解绕荧光分析(FADU)测定DNA链断裂。统计学显著性水平设为alpha = 0.05。与对照组相比,臭氧暴露没有改变TE细胞的产量或活力,但引起气管灌洗中蛋白质含量的增加和DNA链断裂的增加。在3个不同的碱孵育时间下,暴露72 h后TE细胞碱裂解液中留下的DNA量明显低于对照。暴露96 h时,TE细胞碱解液中留下的双链DNA增加,接近暴露24 h时的值。在15℃条件下,三种不同的碱液孵育时间均出现了相同的模式。一个Qd单位估计对应于每个细胞100条链断裂。Qd也被用作O3损伤的指标。与对照相比,无论在15℃下碱孵育时间如何,暴露于1 ppm O3 72 h后,Qd显著增加。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Ozone-induced DNA strand breaks In guinea pig tracheobronchial epithelial cells.

Ozone (O3), the major oxidant of photochemical smog, is thought to be genotoxic and a potential respiratory carcinogen or promoter of carcinogenic processes. Because of oxidative reactions with the mucus in the upper airway, O3 reaction products are able to penetrate into the tracheobronchial epithelial (TE) cells. The carcinogenic effects of O3 on the TE cells are especially of interest since most previous studies have focused on the morphology or permeability changes of tracheas only. Therefore, the objective of this study was to examine the potential O3 genotoxicity in TE cells after an in vivo exposure, using DNA strand breaks as an index. Two-month-old male Dunkin-Hartley guinea pigs, specific pathogen free, 4 in each group, were exposed to 1.0 ppm O3 for 0, 12, 24, 48, 72, or 96 h. Animals exposed to filtered air without O3 exposure were used as controls. After O3 exposure, the trachea with two main bronchi was removed from each animal, and TE cells were isolated and employed for determination of DNA strand breaks by fluorometric analysis of DNA unwinding (FADU). The statistical significance level was set at alpha = .05. Compared with controls, ozone exposure did not alter the TE cell yield or viability, but caused an increase in protein content in tracheal lavage and an increase in DNA strand breaks. The amount of DNA left in the alkali lysate of TE cells found at 72 h exposure was significantly decreased from controls for 3 different alkali incubation times. An increase of the double-stranded DNA left in the alkali lysate of TE cells was observed at 96 h of exposure and approached the value of 24 h of exposure. The same pattern was seen with all 3 different alkali incubation times at 15 degrees C. One Qd unit was estimated to correspond to 100 strand breaks per cell. The Qd was also used as an indicator for O3 damage. Compared to controls, the Qd increases significantly after 1 ppm O3 exposure for 72 h, regardless of the alkali incubation time at 15 degrees C.

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