Luan Shaorong, Yun Xinming, Rao Wenbing, Xiao Ciying, Xu Zhikang, Lang Jialin, Huang Qingchun
Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237, China.
Shanghai Key Lab of Chemical Biology, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
Chem Biol Interact. 2017 Aug 1;273:90-98. doi: 10.1016/j.cbi.2017.06.004. Epub 2017 Jun 7.
Emamectin benzoate (EMB), a novel macrocyclic lactone insecticide, possesses high efficacy and beneficial selective toxicity in agriculture, but so far the EMB-induced cytotoxic action in arthropod insect remains unclear. The present studies were carried out to characterize the property of EMB on the induction of reactive oxygen species (ROS)-mediated DNA damage and apoptosis in Trichoplusia Tn5B1-4 cell model. Following the exposure to EMB at 2.5, 5, 10 or 15 μM, the cells changed to be round, suspended and aggregated, and the decline of cell proliferating ability and cell viability was positively related with the exposure time. Median inhibitory concentration (IC) of EMB on cell viability was 3.72 μM during 72 h exposure. Apoptosis was induced in 29.8% (24 h) and 39.5% (48 h) of the cells by EMB at 15 μM, showing chromatin condensation in nuclei. The content of ROS in the cells increased rapidly as the concentration of EMB increased, and the pre-incubation of the cells with vitamin E significantly reduced the ROS accumulation. In the treatment of 15 μM EMB, the migrated cell nucleus with DNA strand breaks appeared a teardrop, pear-shaped, or large fan-like tail, and 63.1% of γH2AX-positive cells contained more than four foci, accompanying with high expression level of caspase-3 in time-dependent manner, which consequently led to cell apoptotic death. These evidences in ROS-mediated DNA damage and cell apoptosis induced by EMB may be helpful for deep understanding the cytotoxic action of EMB based on cell model.
甲氨基阿维菌素苯甲酸盐（EMB）是一种新型大环内酯类杀虫剂，在农业中具有高效且有益的选择性毒性，但迄今为止，EMB在节肢动物昆虫中引起的细胞毒性作用尚不清楚。本研究旨在表征EMB在粉纹夜蛾Tn5B1-4细胞模型中诱导活性氧（ROS）介导的DNA损伤和细胞凋亡的特性。在2.5、5、10或15μM浓度下暴露于EMB后，细胞变为圆形、悬浮并聚集，细胞增殖能力和细胞活力的下降与暴露时间呈正相关。在72小时暴露期间，EMB对细胞活力的半数抑制浓度（IC）为3.72μM。15μM的EMB诱导29.8%（24小时）和39.5%（48小时）的细胞发生凋亡，细胞核出现染色质凝聚。随着EMB浓度的增加，细胞内ROS含量迅速增加，用维生素E对细胞进行预孵育可显著减少ROS的积累。在15μM EMB处理中，出现DNA链断裂的迁移细胞核呈泪滴状、梨形或大扇形尾巴，63.1%的γH2AX阳性细胞含有四个以上的病灶，并伴随着caspase-3表达水平随时间依赖性升高，最终导致细胞凋亡死亡。这些关于EMB诱导ROS介导的DNA损伤和细胞凋亡的证据可能有助于基于细胞模型深入了解EMB的细胞毒性作用。
