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目的探讨不同作用时间下粒径为25nm的锐钛矿型二氧化钛(TiO2)颗粒对人肺腺癌A549细胞的毒性效应。方法将100μg/ml的纳米TiO2颗粒悬液与A549细胞在37℃、5%CO2的细胞培养箱中分别孵育1h、4h、8h和24h后,收集细胞。通过乳酸脱氢酶(LDH)检测实验检测纳米TiO2颗粒对细胞膜的损伤情况;通过ATP检测实验检测纳米TiO2颗粒对线粒体产生的影响;通过超氧化物歧化酶(SOD)活性检测实验检测纳米TiO2颗粒对细胞的氧化损伤情况;采用透射电子显微镜观察纳米TiO2颗粒引起细胞超微结构的变化;利用CCK-8法检测纳米TiO2颗粒对细胞存活率的影响。结果随着纳米TiO2颗粒作用时间的延长,细胞外液中LDH活性增强,细胞内ATP浓度降低,SOD活性降低,细胞存活率呈现明显下降(P<0.05),且细胞线粒体和内质网出现不同程度的肿胀和扩张。结论纳米TiO2颗粒体外能够引起肺腺癌细胞氧化损伤,抑制细胞生长,且对细胞的毒性效应存在时间依赖性。
Objective To investigate the toxic effects of anatase TiO2 particles with different diameters on the activity of human lung adenocarcinoma A549 cells. Methods After the suspension of 100μg / ml TiO2 nanoparticles were incubated with A549 cells in 37 ℃, 5% CO2 incubator for 1h, 4h, 8h and 24h respectively, the cells were harvested. The effect of nano-TiO2 particles on the mitochondria was detected by ATP assay and the nano-TiO2 particles were detected by the activity test of superoxide dismutase (SOD) The oxidative damage of the cells was observed. The changes of the ultrastructure of the cells were observed by transmission electron microscope. The cell viability was detected by CCK-8 method. Results With the prolongation of the action time, the activity of LDH in extracellular fluid increased, the concentration of ATP in the extracellular fluid decreased, the activity of SOD decreased, the cell viability decreased significantly (P <0.05), and the mitochondria and endoplasmic reticulum were different Degree of swelling and expansion. Conclusion Nano-TiO2 particles can induce oxidative damage of lung adenocarcinoma cells in vitro and inhibit the growth of the cells, and the cytotoxicity of the nanoparticles on the cells is time-dependent.