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In vitro genoprotective and genotoxic effect of nicotine on human leukocytes evaluated by the comet assay

机译:彗星试验评估尼古丁对人白细胞的体外遗传保护和遗传毒性作用

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摘要

The comet assay was used to measure the DNA damage induced in vitro by nicotine in human leukocytes as the extent of DNA migration in the comet head area, tail length, percent DNA in the tail, and Olive tail moment. Samples of whole blood were collected and blood cells were challenged with acute doses of 0.1, 1 and 10 mu M of (-)-nicotine for 60 minutes. We found that nicotine treatment had dose-dependent effects on the level of DNA damage. At 1 and 10 mu M of nicotine, both Olive tail moment and percent DNA in the tail significantly increased (p < 0.001), compared to the control. In the presence of 10 mu M of nicotine, the shortest tail length and the smallest head area were detected. At a concentration of 0.1 mu M, surprisingly, DNA damage detected by the comet assay was lower than in the control, which was proved by the observed significantly (p < 0.001) lower Olive tail moment and percent DNA in the tail as well as larger head area. The results suggest that nicotine, at a reasonably low concentration (0.1 mu M), comparable to those found in the blood of habitual smokers, may have a protective effect, whereas higher doses of nicotine (1 and 10 mu M) are genotoxic. The possible participation of reactive oxygen species in the DNA-damaging potential of nicotine is discussed.
机译:彗星测定法用于测量尼古丁在人白细胞中体外诱导的DNA损伤,如彗星头区域中DNA迁移的程度,尾巴长度,尾巴中的DNA百分比和Olive尾矩。收集全血样品,并用0.1、1和10μM的(-)-尼古丁急性剂量攻击血细胞60分钟。我们发现尼古丁治疗对DNA损伤的水平具有剂量依赖性。与对照组相比,在1和10μM的尼古丁时,橄榄尾矩和尾巴中的DNA百分比均显着增加(p <0.001)。在10μM尼古丁的存在下,检测到最短的尾巴长度和最小的头部面积。令人惊讶的是,在浓度为0.1μM的情况下,彗星试验检测到的DNA损伤低于对照,这被观察到的明显(p <0.001)较低的橄榄色尾矩和尾巴中的DNA百分比以及更大的橄榄证明。头部区域。结果表明,与习惯吸烟者血液中发现的浓度相当的低浓度(0.1μM)的尼古丁可能具有保护作用,而较高剂量的尼古丁(1和10μM)具有遗传毒性。讨论了活性氧可能参与尼古丁破坏DNA的可能性。

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