多壁碳纳米管对神经生长因子诱导的PC12细胞分化的影响

Neurotoxicity of Multi-wall Carbon Nanotubes on PC12 Cells

目的:观察多壁碳纳米管(MWCNTs)对神经生长因子诱导的PC12细胞分化的影响。方法:PC12细胞,随机分为对照组、神经生长因子(NGF)组和NGF+MWCNTs组(n=5),对照组予空白处理,其它2组加入NGF,NGF+MWCNTs组还加入MWCNTs,运用试剂盒CCK-8分别检测5、10、30、60μg/mL MWCNTs孵育48 h后的细胞活性;通过染色观察MWCNTs孵育细胞48 h的细胞状态,免疫荧光检测F-肌动蛋白、β-tubulinⅢ的构象变化。结果:MWCNTs浓度>30μg/mL时有细胞毒性;30μg/mL的MWCNTs孵育细胞48 h,MWCNTs导致F-肌动蛋白、β-tubulinⅢ在细胞边缘表达减弱或凝结成结节样。结论:MWCNTs浓度>30μg/mL,孵育48 h,对细胞有毒性作用,MWCNTs以团状或折叠状存在于细胞内,致细胞凋亡。

Objective： To investigate the effects of Multi-wall carbon nanotubes （MWCNTs） on morphology in PC12 cells. Methods： PC12 cells were cultured and divided into groups control, nerve growth factor （NGF）, and MWCNTs treatments. The control group was given blank treatment. Both groups NGF was added to both groups NGF and MWCNTs, whereas MWCNTs group was treated with additional MWCNTs. CCK-8 assay was used to detect the effect of MWCNTs （5,10,30,60 μg/mL） on cell viability in PC12 cells after 48 h. The morphology of PC12 cells was observed after staining. Immunofluorescence was used to detect the conformational change of F-actin and Beta-tubulin III. Results： MWCNTs had toxic effect on PC12 cells at concentration 〉 30 μg/mL. Im- munofluorescence assay showed that expressions of F-actin and Beta-tubulin IH were decreased in pericellular compartment or condensed into nodular shapes after incubation with 30 txg/mL MWCNTs for 48 h. Conclusion： Incubation with MWCNTs with concentration higher than 30 μg/mL for 48 h inhibits cell proliferation, leading to apoptosis in PC12 cells.