氨甲喋呤对映体对ECV304细胞抑制作用及其机制  

Effects of methotrexate enantiomers on ECV304 cell inhibition and its mechanisms

作　　者：郭立方[1,2] 王荣[1,2] 贾正平[1,2] 施有琴[1,2] 谢华[1] 张娟红[1] 武晓玉[1] 

机构地区：[1]兰州军区兰州总医院药理基地,甘肃兰州730050 [2]兰州大学药学院,甘肃兰州730000

出　　处：《中国药理学通报》2010年第2期213-216,共4页Chinese Pharmacological Bulletin

基　　金：国家科技部重大资助项目(No2008ZXJ-09014-010)

摘　　要：目的探讨氨甲喋呤[(±)MTX]及其对映体[(+)MTX、(-)MTX]对ECV304细胞的生长抑制作用并探讨其机制。方法采用培养的ECV304细胞,应用MTT比色法分析其活性;用光学显微镜观察细胞的形态学变化;碘化丙啶(PI)单染流式细胞术检测细胞周期。结果在0.1～150μmol·L-1范围内,(+)MTX、(-)MTX和(±)MTX作用于ECV304细胞24、48、72h,均抑制细胞ECV304增殖,但抑制强度依次为(+)MTX>(±)MTX>(-)MTX,倒置显微镜观察不同浓度(±)MTX、(+)MTX和(-)MTX作用ECV304细胞不同时间后,出现细胞体积变小、核固缩等形态学改变。用10μmol·L-1的(+)MTX、(-)MTX和(±)MTX作用ECV304细胞48h后,PI单染流式细胞术检测ECV304细胞周期的影响,表明MTX消旋体及单一体干扰ECV304细胞DNA合成。结论(+)MTX和(-)MTX对ECV304细胞的抗增殖作用具有化学结构的立体选择性,(+)MTX的抗ECV304细胞增殖作用明显强于(-)MTX。Aim To investigate the effect of MTX （ibcluded（± ） MTX, （ ＋ ） MTX and（ - ） MTX） on the proliferation of ECV304 cells and to explore its mechanisms. Methods ECV304 cells were cultured. The cell proliferation was determined by MTT. The morphological changes were inspected by inverted microscope. Cell eycle phases were assayed by propidium iodide staining flow eytometry. Results ECV304 cells were treated with（ ＋ ） MTX, （ - ） MTX and（ ＋ ） MTX at 1 150 μmol · L^-1 for 24,48,72 h. The results showed that the proliferation of ECV304 cells was significantly inhibited under different conditions. The order of the inhibited efficacy was （ ＋ ） MTX 〉 （ ± ） MTX 〉 （ - ） MTX. The morphology of ECV304 cells were changedby（ ＋ ） MTX, （ - ） MTX and（ ±） MTX treatment, which included the cell shrinkage, chromatin condensation. After administration of 10 μmol · L^-1 of （ ＋ ） MTX, （ - ） MTX and （ ± ） MTX for 48 h, the cell cycle phases were assayed by propidium iodide staining flow cytometry. The result showed DNA replication was interfered by （ ＋ ） MTX, （ - ） MTX and （ ± ） MTX treatment. Conclusions The proliferation of ECV304 cells has the chiral selective effects by （ ＋ ） MTX and （ - ） MTX treatment, and the inhibition on ECV304 cells proliferation of（ ＋ ） MTX is significantly stronger than that of （ - ）MTX.

关 键 词：氨甲喋呤 对映体药物 ECV304细胞 四甲基偶氮唑蓝比色法 流式细胞术 细胞抑制 机制 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]