Anjos Vanessa A, da Silva Flávio M R, Souza Marta M
Programa de Pós-Graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, 96.203-900, Rio Grande, Rio Grande do Sul, Brazil.
Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, Rio Grande do Sul, Brazil.
Toxicol In Vitro. 2014 Apr;28(3):365-72. doi: 10.1016/j.tiv.2013.11.013. Epub 2013 Dec 8.
Sea anemones are benthic organisms, of low mobility and can be directly affected by water pollution. This work studied the defense mechanisms and DNA damage caused by copper toxicity in cells from the anemone Bunodosoma cangicum. For this, exposure of anemones cells were held, kept in primary culture through explant of podal disk to copper (7.8 and 15.6 μg/L), and the control group, for 6 and 24h. Cytotoxicity was seen through the viability and cell number, MXR phenotype through the accumulation of rhodamine-B, ROS generation by H2DCF-DA and DNA damage by comet assay. The results obtained show that there is a drop in viability and number of cells, especially after exposure of 24h in 15.6 μg/L. There is an induction of the MXR activity only at 7.8 μg/L for 24h. As for ROS, there is an increase in the generation of reactive species in greatest concentration of copper for 6h, and in both for 24h, which leads to oxidative stress, which culminates with a DNA damage. What was evidenced by the increase of the tail size, % DNA presented and moment of tail. Therefore, the copper represents an adversity to the anemones cells, being cytotoxic and genotoxic.
海葵是底栖生物，活动能力低，会直接受到水污染的影响。这项研究了海葵Bunodosoma cangicum细胞中铜毒性引起的防御机制和DNA损伤。为此，将海葵细胞通过足盘外植体进行原代培养，分别暴露于铜（7.8和15.6μg/L）中，对照组则不做处理，处理时间为6小时和24小时。通过细胞活力和细胞数量观察细胞毒性，通过罗丹明-B的积累观察多药耐药（MXR）表型，通过H2DCF-DA检测活性氧（ROS）的产生，通过彗星试验检测DNA损伤。结果表明，细胞活力和数量下降，尤其是在15.6μg/L的铜暴露24小时后。仅在7.8μg/L处理24小时时诱导了MXR活性。至于ROS，在6小时时，铜浓度最高时活性物种的产生增加，在24小时时两种浓度下均增加，这导致氧化应激，最终导致DNA损伤。尾长、呈现的DNA百分比和尾矩的增加证明了这一点。因此，铜对海葵细胞是一种逆境，具有细胞毒性和基因毒性。
