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目的:探讨纳米二氧化硅(SiO2)对人神经母细胞瘤SH-SY5Y细胞凋亡和周期的影响,阐明其作用机制。方法:选取不同浓度粒径为90nm的SiO2颗粒作用于SH-SY5Y细胞,分别为0、3.125、6.250、12.500、25.000、50.000mg·L-1 SiO2组,其中0mg·L-1组为对照组。MTT法检测各组SH-SY5Y细胞存活率;采用AO/EB染色和AnnexinⅤ-FITC/PI法检测各组细胞凋亡率;流式细胞术(FCM)检测细胞周期百分比和细胞中活性氧(ROS)水平。结果:MTT检测,6.250、12.500、25.000和50.000mg·L-1 SiO2组细胞存活率低于对照组(P<0.05),12.500、25.000和50.000mg·L-1 SiO2组细胞存活率低于3.125mg·L-1 SiO2组(P<0.05)。AO/EB染色和Annexin V-FITC/PI法检测,6.250、12.500、25.000和50.000 mg·L-1 SiO2组细胞凋亡率高于对照组(P<0.05),12.500、25.000和50.000 mg·L-1 SiO2组细胞凋亡率高于3.125mg·L-1 SiO2组(P<0.05)。FCM检测,3.125、6.250、12.500、25.000和50.000mg·L-1 SiO2组细胞中ROS水平高于对照组(P<0.05)。25.000和50.000 mg·L-1 SiO2组G0/G1期细胞百分比低于3.125mg·L-1SiO2组和对照组(P<0.05),50.000 mg·L-1 SiO2组G2/M期细胞百分比高于对照组(P<0.05)。结论:纳米SiO2可降低SH-SY5Y细胞的活力,诱导细胞凋亡,增加细胞中ROS水平,可干扰细胞周期进程,对SH-SY5Y细胞的增殖具有抑制作用。
Objective: To investigate the effects of nano-silica (SiO2) on the apoptosis and the cycle of human neuroblastoma SH-SY5Y cells and elucidate its mechanism. Methods: SiO2 particles with different particle size of 90nm were selected to act on SH-SY5Y cells, which were 0, 3.125, 6.250, 12.500, 25.000 and 50.000 mg · L-1 SiO2 respectively. . The survival rate of SH-SY5Y cells in each group was detected by MTT assay. The apoptosis rate of each group was detected by AO / EB staining and AnnexinⅤ-FITC / PI. The percentage of cell cycle and the percentage of reactive oxygen species (ROS) in cells were detected by flow cytometry (FCM) )Level. Results: The viability of cells in 6.250, 12.500, 25.000 and 50.000 mg · L-1 SiO2 groups was lower than that in control group (P <0.05) by MTT assay. The cell viability of 12.500, 25.000 and 50.000 mg · L-1 SiO2 groups was lower than 3.125 mg · L-1 SiO2 group (P <0.05). AO / EB staining and Annexin V-FITC / PI assay showed that the apoptotic rate in the groups of 6.250, 12.500, 25.000 and 50.000 mg · L-1 SiO2 was higher than that of the control group (12.5%, 25.000 and 50.000 mg · L -1 SiO2 group was higher than that of 3.125 mg · L-1 SiO2 group (P <0.05). The levels of ROS in cells of 3.125, 6.250, 12.500, 25.000 and 50.000 mg · L -1 SiO2 groups were higher than that of the control group (P <0.05) by FCM. The percentage of cells in G0 / G1 phase in 25.000 and 50.000 mg · L-1 SiO2 groups was lower than that in 3.125 mg · L-1SiO2 group and the control group (P <0.05). The percentage of G2 / M phase cells in 50.000 mg · L-1 SiO2 group was higher than Control group (P <0.05). Conclusion: Nano-SiO2 can reduce the activity of SH-SY5Y cells, induce apoptosis and increase the level of ROS in cells, which may interfere with the cell cycle progression and inhibit the proliferation of SH-SY5Y cells.