摘要
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 study the damaging effect of nano-neodymium oxide（ Nano-Nd_2O_3） on rat adrenal pheochromocytoma cell PC12. Methods PC12 cells in the logarithmic growth phase were divided into control group and 7 drug treatment groups. The control group was not treated with Nano-Nd_2O_3. The drug treatment groups were treated with 7 different dosing with the final mass concentrations of 3. 13,6. 25,12. 50,25. 00,50. 00,75. 00,100. 00 mg/L of NanoNd_2O_3 for 24 hours,then the follow-up selected experimental concentration were 3. 13,6. 25,12. 50,25. 00,50. 00 and75. 00 mg/L. The CCK-8 essay was used to detect the cell viability. The levels of glutathione（ GSH） and superoxide dismutase（ SOD） in the cells were measured by enzyme-linked immunosorbent assay. The expression of B cell lymphoma-2（ Bcl-2）,Bcl-2 associated X protein（ Bax） and Caspase-3 protein were detected by Western blot. Results The survival rate,GSH level and Bcl-2/Bax ratio of PC12 cells decreased with the increase of the Nano-Nd_2O_3 dose,and showed doseresponse relationship（ P〈0. 01）. The SOD activity of PC12 cells in Nano-Nd_2O_3 group of 12. 50,25. 00,50. 00 and75. 00 mg/L were lower than that of control group（ P〈0. 05）. The ratio of Caspase-3/β-actin in PC12 cells was higher than that in control group（ P〈0. 05）. In the range of 0. 00-25. 00 mg/L of Nano-Nd_2O_3,with the increase of NanoNd_2O_3 dose,the ratio of Caspase-3/β-actin in PC12 cells increased gradually,and showed a dose-response relationship（ P〈0. 01）. Conclusion Nano-Nd_2O_3 can induce oxidative stress in PC12 cells,leading to the decrease of Bcl-2/Bax ratio,activate Caspase-3 expression,and than inhibits cell proliferation in a dose-dependent manner.
出处
《中国职业医学》
CAS
北大核心
2017年第5期552-556,共5页
China Occupational Medicine
基金
国家自然科学基金(81160341)
内蒙古自治区研究生科研创新项目[S201610127(Y01)]
