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目的研究纳米氧化钕(Nano-Nd_2O_3)对大鼠肾上腺嗜铬细胞瘤细胞PC12(以下简称“PC12细胞”)的损伤作用。方法将对数生长期的PC12细胞分为对照组和7个剂量组,对照组不予Nano-Nd_2O_3处理,各剂量组分别予终质量浓度为3.13、6.25、12.50、25.00、50.00、75.00、100.00 mg/L的Nano-Nd_2O_3染毒24 h,选择后续实验剂量。采用CCK-8法检测3.13、6.25、12.50、25.00、50.00、75.00 mg/L Nano-Nd_2O_3组PC12细胞存活率,并采用酶标法测定上述6组细胞的谷胱甘肽(GSH)水平和超氧化物歧化酶(SOD)活力,采用免疫印迹法检测PC12细胞内B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和天冬氨酸特异性半胱氨酸蛋白酶(Caspase)-3蛋白的表达。结果随着Nano-Nd_2O_3染毒剂量的增加,PC12细胞存活率、GSH水平和Bcl-2/Bax比值均下降,均呈剂量-效应关系(P<0.01)。12.50、25.00、50.00和75.00 mg/L Nano-Nd_2O_3组PC12细胞的SOD活力均低于对照组(P<0.05)。各剂量组PC12细胞Caspase-3/β-actin比值均高于对照组(P<0.05),在0.00~25.00 mg/L Nano-Nd_2O_3剂量范围,随着Nano-Nd_2O_3染毒剂量的增加,PC12细胞的Caspase-3/β-actin比值逐渐上升,呈剂量-效应关系(P<0.01)。结论 Nano-Nd_2O_3可导致PC12细胞发生氧化应激,降低Bcl-2/Bax比值,激活Caspase-3表达,从而抑制PC12细胞的增殖,并呈剂量-效应关系。
Objective To investigate the effect of Nano-Nd_2O_3 on the injury of rat adrenal pheochromocytoma cell line PC12 (hereinafter referred to as “PC12 cell”). Methods The logarithmic growth phase of PC12 cells were divided into control group and 7 dose groups, the control group without Nano-Nd_2O_3 treatment, each dose group were given the final concentration of 3.13,6.25,12.50,25.00,50.00,75.00,100.00 mg / L of Nano-Nd_2O_3 exposure 24 h, select the follow-up experimental dose. The cell viabilities of PC12 cells in the Nano-Nd_2O_3 group (3.13,6.25,12.50,25.00,50.00,75.00 mg / L) were determined by CCK-8 assay. The levels of glutathione (GSH) (Bcl-2), Bcl-2 related protein (Bax) and aspartate specific cysteine (Bcl-2) in PC12 cells were detected by Western blot. Expression of Caspase-3 protein. Results With the increase of dose of Nano-Nd_2O_3, the survival rate of PC12 cells, the level of GSH and the ratio of Bcl-2 / Bax were decreased, showing dose-effect relationship (P <0.01). The SOD activity in PC12 cells of 12.50, 25.00, 50.00 and 75.00 mg / L Nano-Nd_2O_3 groups was lower than that of the control group (P <0.05). The ratio of Caspase-3 / β-actin in PC12 cells in each dose group was higher than that in the control group (P <0.05). With the dose of Nano-Nd_2O_3 in the range of 0.00-25.00 mg / L Nano-Nd_2O_3, PC12 cells The ratio of Caspase-3 / β-actin increased gradually with dose-effect relationship (P <0.01). Conclusion Nano-Nd 2 O 3 can induce oxidative stress in PC12 cells, decrease the ratio of Bcl-2 / Bax, and activate Caspase-3 expression, thereby inhibiting the proliferation of PC12 cells in a dose-dependent manner.