| 1-硝基芘和1, 2-萘醌的联合细胞毒性和致DNA损伤 |
| 摘要点击 2372 全文点击 1198 投稿时间：2014-03-28 修订日期：2014-05-30 |
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| 中文关键词 1-硝基芘 1,2-萘醌 细胞毒性 DNA损伤 活性氧自由基 |
| 英文关键词 1-nitropyrene 1,2-naphthoquinone cytotoxicity DNA damage ROS generation |
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| 中文摘要 |
| 以人肺上皮细胞A549为研究对象，运用MTT方法检测1-硝基芘（1-NP）处理后A549的细胞存活率；测定细胞培养液中乳酸脱氢酶（LDH）的漏出率，评价细胞膜损伤；运用彗星实验检测DNA损伤；通过荧光探针的方法测定细胞内产生的活性氧自由基（reactive oxygen species，ROS）. 通过1，2-萘醌（1，2-NQ）预先染毒24 h，再使用1-NP染毒24 h的方法，评估1-NP和1，2-NQ对A549的联合细胞毒性和DNA损伤. 结果表明，1-NP对A549暴露24 h和48 h的半致死浓度（LC50）分别为5.2 μmol·L-1和2.8 μmol·L-1. LC50随着染毒时间的增加而降低，提示暴露时间越长1-NP的细胞毒性越强. A549在1、2、3和4 μmol·L-1浓度的1-NP染毒下，DNA损伤显著增强，ROS水平不断升高，呈现剂量-效应关系（P<0.05）；但LDH漏出率无显著变化. 1，2-NQ（5 μmol·L-1）预染毒A549细胞24 h，能明显减弱1-NP造成的DNA损伤和ROS升高. 结果说明，1，2-NQ预处理可能通过抑制1-NP暴露产生的ROS，从而降低A549的DNA的损伤. |
| 英文摘要 |
| Using human lung epithelial A549 cells, viability was measured by MTT assay after treated with 1-nitropyrene (1-NP); lactate dehydrogenase (LDH) leakage was determined to evaluate the cellular membrane injury; DNA damage was detected with comet assay; reactive oxygen species (ROS) generation was measured with fluorescent probe. The combined toxic effects of 1-NP and 1,2-naphthoquinone (1,2-NQ) on A549 were also evaluated. 1-NP caused a significantly concentration-dependent and time-dependent viability decrease. The LC50 for 24 h and 48 h were 5.2 μmol·L-1 and 2.8 μmol·L-1, respectively. DNA damage and intracellular ROS levels were also increased significantly through a dose-dependent manner after exposure to 1-NP. The LDH leakage were not significantly changed. Compared with the groups treated with 1-NP alone, the viability and LDH leakage was not changed significantly in combined-treated groups with 1-NP and 1,2-NQ. However, the DNA damage and ROS levels were significantly reduced in the combined-treated groups compared with the groups treated with 1-NP alone. These results suggest that 1-NP may mediated the genotoxic and cytotoxic effects through ROS generation, and pretreatment with 1,2-NQ, may inhibit the ROS generation induced by 1-NP, and thereby reducing the DNA damage in A549 cells. |
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