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目的通过研究不同粒径及不同浓度纳米氧化铝颗粒对中国仓鼠肺细胞(CHL细胞)DNA损伤及氧化应激的影响,探索纳米氧化铝颗粒遗传毒性及作用机制。方法选用13、50 nm和10μm 3种粒径的氧化铝颗粒为3个粒径组,每种粒径氧化铝又分别设低剂量(15μg/ml)、中剂量(30μg/ml)、高剂量(60μg/ml)组,另设空白对照组和阳性对照组。染毒24h后处理细胞。用荧光显微镜观察cy 5.5荧光标记纳米颗粒进入细胞的情况。用单细胞琼脂糖凝胶电泳(SCGE)检测DNA损伤,谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)评估氧化应激反应的指标。结果荧光标记的纳米颗粒在细胞核周围分布。SCGE结果显示,与空白对照组相比,各染毒剂量组Olive尾矩有增加趋势,中、高剂量组Olive尾矩均明显增加(P<0.05)。氧化应激指标结果显示,随着染毒剂量的增加各种氧化应激指标明显变化,高剂量组与空白对照组相比,差异有统计学意义(P<0.05)。结论纳米氧化铝颗粒可以引起CHL细胞DNA损伤和氧化应激反应,氧化应激反应可能是导致遗传毒性的机制之一。
OBJECTIVE: To investigate the DNA damage and oxidative stress of Chinese hamster lung cells (CHL cells) with different particle sizes and different concentrations of nano-alumina particles to explore the genetic toxicity and mechanism of nano-alumina particles. Methods Three kinds of alumina particles with three kinds of particle sizes of 13, 50 nm and 10 μm were selected as three particle size groups. Each particle size alumina was divided into low dose (15μg / ml), middle dose (30μg / ml), high dose (60μg / ml) group, another set of blank control group and positive control group. Cells were treated 24h after exposure. Fluorescence microscopy was used to observe cy 5.5 fluorescent-labeled nanoparticles entering cells. The indicators of oxidative stress were evaluated by single cell agarose gel electrophoresis (SCGE) to detect DNA damage, glutathione (GSH), superoxide dismutase (SOD) and malondialdehyde (MDA). Results Fluorescently labeled nanoparticles are distributed around the nucleus. Compared with the blank control group, the Olive tail moment increased in each exposure dose group, while the Olive tail moment in the middle and high dose group increased significantly (P <0.05). The results of oxidative stress indicated that the indicators of oxidative stress changed obviously with the increase of dose, and there was significant difference between high dose group and blank control group (P <0.05). Conclusion Nano-alumina particles can cause DNA damage and oxidative stress in CHL cells. Oxidative stress may be one of the mechanisms leading to genotoxicity.