纳米氧化锌对 HEK293细胞的毒性作用研究

摘要

探讨纳米氧化锌浓度和尺寸效应对HEK293细胞的毒性作用以及其产生毒性的原因。分别用不同浓度的纳米氧化锌（15 nm）和不同尺寸的纳米氧化锌：常规氧化锌（颗粒直径≤1μm）、15 nm氧化锌、30 nm氧化锌及90 nm氧化锌处理HEK293细胞，48 h后，MTT检测纳米氧化锌对HEK293细胞活性的影响，流式细胞仪检测纳米氧化锌对HEK293细胞凋亡的影响，ROS检测试剂盒检测纳米氧化锌对HEK293细胞中ROS含量的影响。结果显示当15 nm氧化锌浓度小于2.0μg／mL时，HEK293细胞活性、凋亡率和ROS水平与对照组相比没有明显差别；但当纳米氧化锌的浓度大于4.0μg／mL时，HEK293细胞活性呈明显下降趋势，细胞凋亡率和ROS含量呈明显的上升趋势，且在同一浓度下，纳米氧化锌尺寸越小，HEK293细胞活性越低，细胞凋亡率和ROS含量越高。该结果表明纳米氧化锌对HEK293细胞的毒性作用呈明显的剂量效应和尺寸效应，纳米氧化锌诱导HEK293细胞产生的大量活性氧（ ROS）可能是导致其产生细胞毒性的主要原因之一。%Abstract This experiment explored the effects of concentration and size of ZnO nanoparticles ( ZnO NPs) on HEK293 cells and re-search its mechanism of toxicity .In this experiment, three different sizes ZnO NPs including 15 nm-sized ZnO NPs, 30 nm-sized ZnO NPs, 90 nm-sized ZnO NPs and normal ZnO to treat HEK293 cells were used.After 48 hours, the cell proliferation was detected by MTT test, then a cell cytometry was made to investigate the rate of apoptosis .Lastly, the change of ROS level was analyzed by kit . There was no noticeable changes on HEK 293 cells treated with 15 nm-sized ZnO NPs below 2.0 μg/mL concentration .While above 4.0 μg/mL, the proliferation of HEK293 cells was reduced significantly .However the rate of apoptosis and the level of ROS increased obviously.With the decrease of the size of ZnO NPs , the rate of apoptosis and the level of ROS increased , but the proliferation of cell gradually reduced in HEK293 cells.The results suggested that cytotoxicity of ZnO NPs on HEK 293 cells was related to its concentration and size.Increasing of ROS induced by ZnO NPs may be one important element of cytotoxicity .