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[目的]比较不同粒径(15、30、100 nm)纳米二氧化硅(nano-SiO2)和常规二氧化硅(Micro-SiO2)对人皮肤表皮细胞(HaCaT)生长的抑制作用及凋亡的影响。[方法]采用不同浓度(2.5、5、10μg/ml)不同粒径(15、30、100 nm)的nano-SiO2和10μg/ml的Micro-SiO2(1～5μm)染毒体外培养的HaCaT细胞24 h,同时设溶剂对照组(nano-SiO2和Micro-SiO2的分散液染毒组)。采用细胞计数试剂盒(CCK-8)法检测HaCaT细胞暴露于不同浓度、不同粒径的nano-SiO2后的细胞活力,用Annexinⅴ-PI双染法和Hoechest33342染色法测定细胞凋亡情况;使用2’,7’-二乙酰二氯荧光素荧光探针检测细胞内活性活性氧(ROS)水平。[结果]不同粒径的nano-SiO2纳米对HaCaT细胞的半数抑制浓度分别为(19.4±1.3)、(27.7±1.5)、(35.9±1.6)μg/ml。当染毒浓度固定时,随着nano-SiO2粒径的减小,凋亡率逐渐增加。当粒径固定时,胞内ROS的水平也随着nano-SiO2浓度的增大而增高。相关分析表明,细胞存活率和凋亡率与胞内活性氧的水平的相关性分别为-0.952和0.898(P<0.01)。[结论]nano-SiO2能抑制HaCaT细胞生长并可诱导其凋亡,这种现象的发生可能与胞内产生的活性氧水平有关。
[Objective] To compare the inhibitory effects of different particle sizes (15, 30, 100 nm) nano-SiO2 and conventional silica on human skin epidermal cells (HaCaT) influences. [Method] HaCaT cells cultured in vitro with different concentrations (2.5, 5, 10μg / ml) of nano-SiO2 and 10μg / ml Micro-SiO2 24 h, while the solvent control group (nano-SiO2 and Micro-SiO2 dispersion liquid exposure group). The cell viability of HaCaT cells exposed to nano-SiO2 with different concentrations and different particle sizes was detected by the cell counting kit (CCK-8) method. The apoptosis of HaCaT cells was detected by Annexin ⅴ-PI double staining and Hoechest33342 staining. ’, 7’-diacetyl dichlorofluorescein fluorescence probe to detect intracellular reactive oxygen species (ROS) levels. [Results] The median inhibitory concentrations of nano-SiO2 nanoparticles with different particle sizes on HaCaT cells were (19.4 ± 1.3), (27.7 ± 1.5) and (35.9 ± 1.6) μg / ml, respectively. When the exposure concentration was fixed, with the decrease of nano-SiO2 particle size, the apoptosis rate increased gradually. When the particle size was fixed, the level of intracellular ROS also increased with the increase of nano-SiO2 concentration. Correlation analysis showed that the correlation between cell viability and apoptosis rate and intracellular reactive oxygen species levels were -0.952 and 0.898, respectively (P <0.01). [Conclusion] nano-SiO2 can inhibit the growth of HaCaT cells and induce its apoptosis. The occurrence of this phenomenon may be related to the intracellular reactive oxygen species level.