摘要
目的研究苯及其代谢产物氢醌对外周血人淋巴细胞周期阻滞与凋亡的影响,探讨苯的细胞毒性作用机制。方法离体培养人淋巴细胞24 h后加S9液,设置苯低、中、高浓度(0.25、3.5、50μmol/L)和氢醌低、中、高浓度(50、150、450μmol/L)的染毒组,另设空白对照组和溶剂对照组,采用四甲基偶氮唑蓝比色法检测细胞相对存活率,流式细胞术检测细胞周期和凋亡的分布状况,荧光检测细胞存活率的含量,单细胞凝胶电泳技术检测DNA断裂。结果苯与氢醌剂量依赖性降低人淋巴细胞存活率,诱导人淋巴细胞阻滞于S+G2/M期,并明显促凋亡且随着染毒浓度升高细胞内活性氧含量增加,与对照组比较,差异有统计学意义(P<0.05),苯与氢醌高浓度组彗星尾长分别为(26.45±7.96)、(30.28±6.07)μm,均明显高于对照组(P<0.01)。结论苯及其代谢物氢醌在体外可导致人淋巴细胞存活率降低,细胞周期紊乱,其机制与细胞内活性氧产生及DNA-蛋白质损伤有关。
Objective To study the effects of benzene and the benzene metabolite hydroquinone on the cell cycle and apoptosis of peripheral blood lymphocytes,and to explore the molecular mechanisms underlying benzene-induced cytoxicity damage. Methods Human lymphatic cells were isolated,cultivated and then divided into three groups including of low,moderate,and high benzene exposure（0.25,3.5,50 μmol/L） and three groups of low,moderate,and high hydroquinone exposure（50,150,450 μmol/L）.One solvent control and one blank control group were also set up.After the treatment,we assayed the growth arrest of lymphocytes induced by benzene and hydroquinone by methyl thiazolyl tetrazolium（MTT）test.Flow cytometry was applied to detect cell cycle and apoptosis rate.2,7-dichlorodihydro fluorescein diacetate（DCFH-DA） assay was used to detect reactive oxygen species（ROS） contents.Lymphocytes′DNA fragment was detected with single cell gel electrophoresis technology. Results The results showed that both benzene and hydroquinone decreased cell viability in a dose dependent manner,along with induction of S phase and G2/M phase arrest and increased late apoptosis.Meanwhile,benzene and hydroquinone induced accumulation of intracellular ROS in a dose-effect relationship.All data showed significant difference compared to control groups（P0.05）.The mean tail lengths were 26.45±7.96 and 30.28±6.07 μm for groups of high levels of benzene and hydroquinone.The results revealed a significantly higher level of DNA damage in all concentration groups compared to control groups with a dose-effect relationship. Conclusion Benzene and its′ metabolite hydroquinone could induce decreased lymphocytes viability,dysfunction of cell cycle and programmed cell death.The results indicate that the overproduction of ROS and DNA lesion are likely to contribute to benzene-induced cytogenetic changes.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2011年第11期1426-1428,共3页
Chinese Journal of Public Health
基金
广西科技厅资助课题(桂科攻0472002-12)
关键词
苯
氢醌
细胞周期
细胞凋亡
氧化损伤
DNA损伤
benzene
hydroquinone
cell cycle
cell apoptosis
oxidative damage
DNA lesion
