论文部分内容阅读
[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1~3d的SD大鼠神经元细胞,培养5d。以BaP同时加入S9对细胞染毒,染毒组浓度分别为10、20、40μmol/L,并设空白对照组和溶剂对照组,继续培养48h。以20μmol/L BaP染毒,分别于染毒0、6、12、24、48h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、40μmol/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0h组相比,染毒48h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,40μmol/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0h组相比,染毒48h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。
[Objective] To explore the mechanism of BaP-induced neuronal apoptosis by studying DNA damage and neuronal cell cycle changes in neurons after benzo [a] pyrene (BaP) exposure. [Method] The neonatal rat neurons of 1 ~ 3d SD rats were cultured for 5 days. The cells were exposed to BaP while S9 cells were exposed to the concentrations of 10, 20, and 40μmol / L, respectively. Blank control group and solvent control group were established and cultured for 48h. The cells were treated with 20μmol / L BaP, and treated at 0, 6, 12, 24, 48h after exposure respectively. DNA damage was detected by single cell agarose gel electrophoresis (SCGE), and cell cycle changes were detected by flow cytometry. [Results] Compared with the blank control group and the solvent control group, the results of SCGE showed that the Olive tail moment, the tail DNA content and the tail length of neuronal cells in 20 and 40 μmol / L groups increased significantly and the content of DNA in head decreased significantly Statistical significance (P <0.05). Compared with 0h group, Olive tail moment, tail DNA content and tail length of neuronal cells in 48h exposure group were significantly increased, and the content of DNA in head decreased significantly, the differences were statistically significant (P <0.05). Flow cytometry results showed that compared with the blank control group, the percentage of neurons in G1 phase and S phase in 40μmol / L treatment group was significantly different (P <0.05); Compared with 0h group, 48h The percentage of neurons in G1 phase and S phase increased obviously, with statistical significance (P <0.05). [Conclusion] BaP can cause DNA breakage in nerve cells and induce cell cycle restart.