NNK对NCTC 1469细胞毒性作用的研究

发布时间：2019-07-03 13:24

【摘要】：研究4-(甲基亚硝氨基)-1-(3-吡啶基)-1-丁酮(NNK)对小鼠肝细胞NCTC 1469细胞的毒性作用。利用MTT比色法检测5个不同NNK浓度(0.1、0.2、0.3、0.4、0.5 mg/m L)对细胞的毒性作用;倒置相差显微镜观察细胞的形态变化;流式细胞仪检测细胞凋亡率;实时荧光定量PCR检测凋亡基因Bcl-2和Bax的表达。MTT显示,NNK能降低NCTC 1469细胞存活率,并呈剂量和时间依赖性。镜检结果可见细胞皱缩,密度降低;流式细胞仪检测显示,0.1、0.2、0.3、0.4、0.5 mg/m L的NNK作用24 h后凋亡率分别为(16.79±2.01)%、(26.87±1.67)%、(41.78±3.19)%、(50.89±3.94)%和(65.86±4.54)%,显著高于对照组(7.46±1.58)%;荧光定量PCR结果显示,随着NNK浓度的增加,Bcl-2 m RNA的表达减少,Bax m RNA的表达逐渐上升。NNK能够降低NCTC 1469细胞存活率并诱导其凋亡,作用呈剂量和时间依赖性。
[Abstract]:The toxic effect of 4-(methyl nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on mouse hepatocytes NCTC 1469 cells was studied. The toxic effects of five different concentrations of NNK (0.1, 0.2, 0.4, 0.4, 0.5 mg/m / L) were detected by MTT colorimetric assay, the morphological changes were observed by inverted phase contrast microscope, the apoptosis rate was detected by flow cytometry, and the expression of apoptosis genes Bcl-2 and Bax was detected by real-time fluorescence quantitative PCR. PCR showed that NNK could decrease the survival rate of NCTC 1469 cells in a dose-and time-dependent manner. The results of microscopic examination showed that the apoptosis rate of cells was (16.79 卤2.01)%, (26.87 卤1.67)%, (41.78 卤3.19)%, (50.89 卤3.94)% and (65.86 卤4.54)%, respectively, which was significantly higher than that of the control group (7.46 卤1.58)%, and the apoptosis rate was (16.79 卤2.01)%, (26.87 卤1.67)%, (41.78 卤3.19)%, (50.89 卤3.94)% and (65.86 卤4.54)%, respectively. The results of fluorescence quantitative PCR showed that the expression of Bcl-2 m RNA decreased and the expression of, Bax m RNA increased gradually with the increase of NNK concentration. NNK could decrease the survival rate and induce apoptosis of NCTC 1469 cells in a dose-and time-dependent manner.
【作者单位】：郑州轻工业学院食品与生物工程学院;
【基金】：郑州市科技局科技攻关项目(121PPTGG362-16) 河南省教育厅青年骨干教师项目(2012GG-117)
【分类号】：R114

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