氯丙咪嗪对脑胶质瘤U251细胞活性的抑制作用及机制  

Inhibition effects of chlorimipramine on human brain U251 glioma cells and its potential mechanism

作　　者：朱勋[1] 王菁哲[1] 陈品 王存祖[1] 许慧中[3] 欧阳琦[1] 

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]苏北人民医院神经外科,江苏扬州215001 [3]东台市人民医院神经外科,江苏东台224200

出　　处：《江苏大学学报（医学版）》2016年第2期132-136,共5页Journal of Jiangsu University：Medicine Edition

摘　　要：目的:探讨氯丙咪嗪促进人脑胶质瘤U251细胞凋亡的可能机制。方法:用甲基噻唑基四唑(MTT)比色法检测不同浓度(228、114、57、28.5、14.25μmol/L)的氯丙咪嗪对U251细胞增殖活性的影响,选取57、28.5μmol/L浓度组进行台盼蓝染色、细胞凋亡染色、流式细胞仪检测,采用蛋白质印迹法和免疫荧光染色法检测细胞中转位分子蛋白(translocator protein,TSPO)的表达,2',7'-二氢二氯荧光黄双乙酸钠(DCFH-DA)探针法检测细胞内活性氧的水平,荧光素酶法检测细胞内ATP含量。结果:氯丙咪嗪组细胞死亡率及凋亡率明显增加,且一定范围内与药物浓度相关;与对照组相比,氯丙咪嗪组TSPO的表达明显增加(P<0.05),活性氧生成明显增加,而细胞内ATP含量明显降低(P<0.05)。结论:氯丙咪嗪可能通过增加细胞内TSPO的表达及活性氧水平,降低ATP含量而促进U251细胞凋亡。Objective： To explore the possible mechanism of chlorimipramine on human brain glioma U251 cell apoptosis. Methods： Using methyl thiazolyl tetrazolium （MTF） colorimetry with different concentrations of ehlorimipramine to test the effects of chlorimipramine on human brain U251 glioma cells; based on the re- suits, selecting two group of different concentrations for the detection of relevant indicators, including death count of tlypan blue staining cell, apoptosis staining, the detection of flow cytometry. Additionally, Western blotting and immumofluorescence assay were conducted to examine the expression level of transloeator protein （TSPO）. DCFH-DA probe was performed to examine the level of reactive oxygen species （ROS） and lucifer- ase assay was applied to investigate cell mitochondrial membrane potential. Results： Based on the testing re- sults of MTT, selecting chlorimipramine 57, 28.5 μmol/L, the concentrations of which have statistically sig- nificant differences; then putting them into the following tests： trypan blue staining, cell apoptosis staining, and flow cytometry. All the testing results confirmed that chlorimipramine had significantly inhibiting effects on human brain U251 glioma cells, and with a certain degree, the effects were associated with the drug con- centration. Additionally, comparing with the control group, Western blotting analysis and immumofluores- cence demonstrated that the expression level of TSPO was significantly increased in the groups of chlorimipra- mine （P 〈 0.05 ）. The level of ROS was significantly higher, but ATP quantity decreased significantly （ P 〈 0.05 ）. Conclusion： Chlorimipramine could significantly inhibit human U251 glioma cell growth, promote apop- tosis by increasing the expression level of TSPO, ROS and decreasing ATP quantity. Chlorimipramine has good anti-glioma effects, and could be used as a kind of candidate drug for the treatment of glioma chemotherapy.

关 键 词：神经胶质瘤 氯丙咪嗪 增殖 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]