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目的探究地表水中有机提取物对人B淋巴细胞DNA的损伤效应。方法于2010年6—8月,采集某江流域18个采样点水样并提取有机物。将处于对数生长期的人B淋巴母细胞悬液分别暴露于0(对照)、250、500、1 000 ml/ml水样有机提取物培养24 h;采用CCK-8活细胞计数试剂盒和彗星试验分别研究水样有机提取物对细胞存活率的影响及其DNA损伤效应,并检测细胞内活性氧自由基(ROS)含量的变化,以评价水样有机提取物的氧化损伤效应。结果与对照组比较,除250 ml/ml采样点5、11、12水样外,各采样点水样有机提取物在各暴露剂量下对人B淋巴母细胞的存活率均较低,差异有统计学意义(P<0.01);且随着各采样点水样有机提取物暴露剂量的升高,人B淋巴细胞的存活率呈下降趋势。与对照组相比,采样点4、5、6水样有机提取物在250、500、1 000 ml/ml剂量下对人B淋巴母细胞彗星尾部DNA含量无明显影响,采样点9、11、12和15水样有机提取物仅在1 000 ml/ml剂量下导致人B淋巴母细胞尾部DNA含量增加,差异有统计学意义(P<0.05,P<0.01);其余各采样点水样有机提取物在500、1 000 ml/ml剂量下均可导致人B淋巴母细胞彗星尾部DNA含量增加,差异有统计学意义(P<0.05,P<0.01)。与对照组相比,采样点16水样有机提取物在500 ml/ml剂量下对人B淋巴细胞内ROS含量无显著影响(P>0.05),其余各采样点水样有机提取物在250~1 000 ml/ml剂量范围内均可显著诱导人B淋巴细胞内ROS的产生,差异有统计学意义(P<0.01);除采样点15外,随着各采样点水样有机提取物暴露剂量的升高,人B淋巴细胞中ROS的含量均呈上升趋势。结论该流域江水中有机提取物对人B淋巴细胞DNA有显著的损伤效应,可能与细胞内ROS含量增加有关。
Objective To investigate the DNA damage effects of organic extracts from surface water on human B lymphocytes. Methods From June to August in 2010, 18 samples of water samples from a river basin were collected and the organic matter was extracted. Human B lymphoblastoid cell suspensions in logarithmic growth phase were exposed to 0 (control), 250, 500, 1000 ml / ml aqueous organic extracts were cultured for 24 h; CCK-8 live cell counting kit and The comet assay was used to study the effect of water-based organic extract on cell viability and its DNA damage effect, and to detect the change of reactive oxygen species (ROS) content in cells to evaluate the oxidative damage effect of organic extracts. Results Compared with the control group, except the 5, 11, and 12 water samples at the 250 ml / ml sampling points, the survival rates of the human B lymphoblasts at various exposure doses were lower for all water samples, Statistical significance (P <0.01). And with the increase of exposure dose of organic extract at each sampling point, the survival rate of human B lymphocytes decreased. Compared with the control group, the sampling points 4, 5 and 6 water samples had no significant effect on DNA content of comet tail of human B lymphoblasts at the doses of 250, 500 and 1000 ml / ml. The sampling points 9, 11, 12 and 15 water samples lead to an increase of tail DNA content of human B lymphoblasts at a dosage of 1 000 ml / ml, with statistical significance (P <0.05, P <0.01) Extracts at 500 and 1 000 ml / ml all resulted in an increase in DNA content in comet tail of human B lymphoblasts, with statistical significance (P <0.05, P <0.01). Compared with the control group, the sampling point 16 water-like organic extract had no significant effect on ROS content in human B lymphocytes at 500 ml / ml (P> 0.05) 1 000 ml / ml dose range can significantly induce ROS production in human B lymphocytes, the difference was statistically significant (P <0.01); except for the sampling point 15, with the sampling point of water-based organic extract exposure dose The level of ROS in human B lymphocytes increased. Conclusion Organic extracts from the river water have a significant damage effect on DNA of human B lymphocytes, which may be related to the increase of intracellular ROS content.