纳米硫化镉对A549细胞的损伤研究

研究纳米硫化镉(Nano-Cd S)材料对肺癌细胞系A549的毒性及氧化损伤作用。培养A549细胞,经传代后接种于6孔板中,每孔2 m L完全培养基,接种次日进行染毒。用直径20~30 nm、长度80~100 nm的Nano-Cd S进行染毒,染毒浓度分别为0、5、10、20、40和80 mg·L~(-1)。染毒24 h后用MTT检测细胞存活率,以存活率在80%左右的浓度为后续实验染毒浓度。应用流式细胞技术,用荧光探针法检测A549细胞的活性氧(reactive oxygen species,ROS)含量,PI-Annexin-V法检测细胞凋亡情况;用试剂盒检测细胞中超氧化物岐化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性以及丙二醛(malondialdehyde,MDA)含量,判断细胞氧化损伤情况。不同浓度Nano-Cd S处理细胞24 h之后,细胞存活率随剂量的增加而下降,浓度为10、20、40和80μg·L~(-1)时,存活率分别为(88.71%±0.80%)、(81.93%±3.06%)、(75.23%±1.13%)和(70.66%±5.63%),且各组间差异均具有统计学意义(P0.05)。以浓度为10和20 mg·L~(-1)的Nano-Cd S染毒24 h后,胞内ROS含量和细胞凋亡率随染毒剂量的增加而增加(P0.05);浓度为10 mg·L~(-1)时,细胞凋亡率为(6.26%±0.44%)。与对照相比,各染毒组SOD和CAT活性和MDA含量升高,20 mg·L~(-1)染毒组SOD和CAT活性和MDA含量高于10 mg·L~(-1)染毒组(P0.05)。研究表明,纳米硫化镉能引起A549细胞的氧化损伤和细胞凋亡,具有明显的细胞毒性。

Damage of A549 Cells Induced by Nanometer Cadmium Sulfide Exposure

This paper aims to investigate the oxidative damage of the lung cancer cell line A549 following nanometer cadmium sulfide (Nano-CdS) exposure.A549 cells were cultured in 6-well plates with 2 mL complete medium and then treated with Nano-CdS for 24 h, whose diameters ranged from 20 to 30 nm and lengths were from 80 to 100 nm.The doses of Nano-CdS were 0, 5, 10, 20, 40 and 80 mg·L-1 respectively.MTT colorimetric assay was used to detect the cell survival rate, and the dose resulting in 80% cell survival rate was used as subsequent exposure dose.Flow cytometry was applied to detect the reactive oxygen species (ROS) contents of A549.The cell apoptosis was assayed by using PI-Annexin-V.The activities of SOD and CAT, the content of MDA were detected with the kits.After treatment with different concentrations of Nano-CdS for 24 h, the survival rates of cells were (88.71%±0.80%), (81.93%±3.06%), (75.23%±1.13%) and (70.66%±5.63%) respectively at doses of 10, 20, 40 and 80 mg·L-1, showing a signficant decrease (P <0.05) with Nano-CdS dose increase.Intracellular ROS contents and apoptosis rate of cells increased significantly after being treated with 10 or 20 mg·L-1 Nano-CdS, compared with the control (P <0.05).The apoptosis rates were (6.26%±0.44%) and (8.94%±1.38%) at 10 and 20 mg·L-1 Nano-CdS groups.Compared with control, the activities of SOD and CAT, and the content of MDA all increased.The activities of SOD and CAT, and the content of MDA in 20 mg·L-1 exposure group were significantly higher compared to 10 mg·L-1 exposure group (P<0.05).It is indicated that Nano-CdS exposure can cause oxidative damage and apoptosis in A549 cells and then result in cytotoxicity.