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目的:研究苯致人外周血单个核细胞(PBMCs)周期阻滞与凋亡,探讨苯对PBMC DNA氧化损伤效应的影响。方法:离体培养24 h后加S9液,设置苯低、中、高浓度组(0.5,5,50μmol/L)和乙醇溶剂对照组,染毒处理24 h后,采用四唑盐比色法检测PBMCs相对存活率;流式细胞术检测细胞周期构成比及凋亡率;DCFH-DA荧光探针检测活性氧(ROS)含量、黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活力;硫代巴比妥酸TBA比色法测定丙二醛(MDA)含量;酶联免疫吸附法(ELISA)测定细胞中谷胱甘肽(GSH)含量;SCGE检测DNA断裂;微核实验检测染色体畸变。结果:与对照组相比,0.5μmol/L苯染毒组细胞存活率明显下降(P<0.05),G0/G1期细胞百分比明显减少(P<0.05),S期和G2/M期百分比明显增加(P<0.05),且细胞凋亡率明显上升(P<0.05),细胞内ROS、MDA、微核率、彗星率和彗星尾长呈剂量依赖性增加,差异均具有统计学意义(P<0.05),而SOD和GSH呈剂量依赖性减少(P<0.05)。结论:苯可以导致PBMCs细胞凋亡和S+G2/M期阻滞,增加细胞脂质过氧化作用,改变细胞内氧化还原状态,诱导DNA氧化损伤。
OBJECTIVE: To study the cycle arrest and apoptosis of human peripheral blood mononuclear cells (PBMCs) induced by benzene and to explore the effect of benzene on DNA oxidative damage induced by PBMC. Methods: S9 liquid was prepared after 24 h culture in vitro, and the control group was treated with low, medium and high concentration of benzene (0.5, 5, 50 μmol / L) and alcohol solvent for 24 h. The relative survival rate of PBMCs was detected by flow cytometry. The cell cycle composition ratio and apoptosis rate were detected by flow cytometry. The content of reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe, and the superoxide dismutase (SOD) activity was measured by xanthine oxidase method. Thiobarbituric acid TBA colorimetric method was used to determine the content of malondialdehyde (MDA); glutathione (GSH) content was measured by enzyme-linked immunosorbent assay (ELISA); DNA fragmentation was detected by SCGE; Results: Compared with the control group, the cell viability in the group treated with 0.5 μmol / L Benzene was significantly decreased (P <0.05), the percentage of cells in the G0 / G1 phase was significantly decreased (P <0.05), and the percentage of S phase and G2 / M phase was significantly (P <0.05), and the apoptosis rate increased significantly (P <0.05). The intracellular ROS, MDA, micronucleus rate, comet rate and comet tail length increased in a dose-dependent manner, the differences were statistically significant (P <0.05), while SOD and GSH decreased in a dose-dependent manner (P <0.05). CONCLUSION: Benzene can lead to cell apoptosis and S + G2 / M arrest in PBMCs, increase lipid peroxidation, change intracellular redox status and induce DNA oxidative damage.