席华星, 聂继盛, 牛侨. 苯并[a]芘对神经细胞DNA损伤及细胞周期的影响[J]. 环境与职业医学, 2012, 29(10): 620-623,628.
引用本文: 席华星, 聂继盛, 牛侨. 苯并[a]芘对神经细胞DNA损伤及细胞周期的影响[J]. 环境与职业医学, 2012, 29(10): 620-623,628.
XI Hua-xing , NIE Ji-sheng , NIU Qiao . Benzo[a]pyrene Induced Neuronal Cell DNA Damage and Cell Cycle Changes in Vitro[J]. Journal of Environmental and Occupational Medicine, 2012, 29(10): 620-623,628.
Citation: XI Hua-xing , NIE Ji-sheng , NIU Qiao . Benzo[a]pyrene Induced Neuronal Cell DNA Damage and Cell Cycle Changes in Vitro[J]. Journal of Environmental and Occupational Medicine, 2012, 29(10): 620-623,628.

苯并a芘对神经细胞DNA损伤及细胞周期的影响

Benzoapyrene Induced Neuronal Cell DNA Damage and Cell Cycle Changes in Vitro

  • 摘要: 目的 通过研究苯并a芘(BaP)染毒后神经细胞DNA损伤和神经元细胞周期改变,探索BaP致神经细胞凋亡的机制。

    方法 采用新生1~3 d的SD大鼠神经元细胞,培养5 d。以BaP同时加入S9对细胞染毒,染毒组浓度分别为10、20、40 μmol/L,并设空白对照组和溶剂对照组,继续培养48 h。以20 μmol/L BaP染毒,分别于染毒0、6、12、24、48 h后处理细胞。用单细胞琼脂糖凝胶电泳(SCGE)检测DNA损伤,采用流式细胞仪检测细胞周期变化情况。

    结果 SCGE结果显示,与空白对照组和溶剂对照组相比, 20、40 μmol/L染毒组神经细胞Olive尾矩、尾DNA含量、尾长均明显增加,头DNA含量明显降低,且有统计学意义(P<0.05)。与0 h组相比,染毒48 h组神经细胞Olive尾矩、尾DNA含量、尾长均有明显增加,头DNA含量明显降低,差异均有统计学意义(P<0.05)。流式细胞仪检测结果显示,与空白对照组相比, 40 μmol/L染毒组G1期和S期神经细胞百分数差异有统计学意义(P<0.05);与0 h组相比,染毒48 h组G1期和S期神经细胞百分数明显增加,且差异有统计学意义(P<0.05)。

    结论 BaP可引起神经细胞DNA断裂,诱发细胞周期重启。

     

    Abstract: Objective To study the mechanism of neuronal apoptosis induced by on DNA damage and cell cycle changes in nerve cells after exposed to benzoapyrene (BaP).

    Methods The neurons from cerebral cortex of neonatal SD rats (1-3 d) were incubated for 5 days.Then the cells were divided into two lots.The first lot was separated into 5 groups,namely blank control,low-dose,medium-dose,high-dose and solvent control groups,and intoxicated by BaP with S9 added at final concentrations of 0,10,20,40μmol/L and dimethyl sulfoxide respectively,then incubated for 48 hours.The second lot was separated into 5 groups,treated with 20μmol/L BaP and cultured for 0 h,6 h,12 h,24 h,and 48 h respectively.DNA damage was detected by single cell gel electrophoresis (SCGE) assay and cell cycle changes by flow cytometry.

    Results SCGE results suggested that compared with the blank control group and the solvent control group,the Olive tail moment,tail DNA percent and tail length of the 20 and 40 μmol/L groups were significantly increased,and the head DNA percent of both groups were significantly decreased (P<0.05).Compared with the 0 h group,the Olive tail moment,tail DNA percent and tail length of the 48 h group were increased notably,and the head DNA percent was reduced (P<0.05).The neuron percentages of G1 period and S period were significantly higher in the 48 h group than in the 0 h group (P<0.05).

    Conclusion BaP could induce neural cell DNA cleavage and cell cycle re-entry.

     

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