不同粒径纳米氧化铝对体外培养神经细胞凋亡的影响
In Vitro Study on the Effects of Nano-Aluminum Oxide Particles with Various Sizes on Apoptosis of Neuron Cells
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摘要:
目的 体外检测不同粒径纳米氧化铝颗粒对昆明乳鼠神经细胞凋亡的影响。 方法 透射电镜观察纳米氧化铝形态与尺寸, 以 10 nm、50 nm、10 μm Al2O3同剂量对体外培养的昆明乳鼠进行染毒, 运用细胞计数试剂盒(cell counting kit, CCK-8)试剂检测细胞活力, 用吖啶橙/溴化乙锭(acridine orange/ethidium bromide, AO/EB)双重染色法观察细胞死亡的形态学, 采用 Rhodamine123染色方法检测线粒体膜电位变化情况, 间接反映早期凋亡变化, 应用流式细胞仪测定各染毒组细胞凋亡。 结果 纳米粒子超声后, 静置前后相比, 电镜下可见纳米粒子均匀分布, 且粒径小于100 nm符合实验要求; AO/EB染色后, 细胞形态学发生明显改变; 罗丹明测线粒体膜电位, 随着纳米氧化铝粒径的减小,膜电位逐渐降低; 应用流式细胞术检测凋亡, 随着纳米氧化铝粒径的减小, 凋亡率逐渐增加。 结论 氧化铝颗粒能够引起原代培养的神经细胞凋亡, 且粒径能够影响细胞凋亡率。 Abstract:Objective To observe the apoptosis of cultured neurons from neonatal mice induced by nano-aluminum oxide (Al2O3) particles with different sizes in vitro. Methods Cultured neurons of neonatal mice were treated with nanoaluminum oxide at same doses but with different sizes. The morphological changes of neurons were observed under trans mission electron microscopy with acridine orange (AO)/ethidium bromide (EB) double staining. The cell viability was examined with cell counting kit (CCK-8) assay. The collapses of mitochondrial trans membrane potential (ΔΨm) were measured using Rhodamine123 staining to detect the undergoing apoptosis and the apoptosis rates were checked by flow cytometry. Results After ultrasonic for 10 min, the nanoparticles were uniformly dispered with their particle sizes less than 100 nm under an electron microscopy both prior to and after standing. After being exposed to nano-alumina, the morphology of neurons changed obviously. With the decrease of nanoparticle size, the apoptosis rates and the ΔΨm loss increased significantly. Conclusion Nanoalumina particles may induce apoptosis in primarily cultured mice neurons. In addition, the particle size of nano-alumina may influence the apoptosis rate.
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