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大鼠原代肺细胞体外非预定DNA合成试验的特性分析。

Characterization of the in vitro unscheduled DNA synthesis assay in primary lung cells of the rat.

作者信息

Whong W Z, Stewart J D, Ong T

机构信息

Division of Respiratory Disease Studies, National Institute for Occupational Safety and Health, Morgantown, WV 26505.

出版信息

Mutat Res. 1991 Jan;262(1):51-5. doi: 10.1016/0165-7992(91)90106-e.

Abstract

The in vitro unscheduled DNA synthesis (UDS) assay has been evaluated in rat primary lung cells with known genotoxicants. The autoradiographic method was employed to detect UDS in both alveolar macrophages and primary pulmonary cells. Data of a time course study revealed that a high radioactive labeling of DNA repair was achieved after a 16-h incubation with [3H]thymidine. Coupled with low serum (1%), hydroxyurea at the concentration of 20 mM inhibited regular DNA synthesis in primary lung cells in a satisfactory manner (81-88% inhibition). With this protocol, a dose-related increase in UDS was induced by N-methyl-N'-nitro-N-nitrosoguanidine and 2-aminoanthracene in both rat alveolar macrophages and primary lung cells. The results suggest that primary rat lung cells in culture possess DNA-repair ability and that the UDS assay may be useful for assessing the pulmonary genotoxic effect of chemicals in this cell system.

摘要

已使用已知的基因毒性剂在大鼠原代肺细胞中评估了体外非预定DNA合成(UDS)试验。采用放射自显影法检测肺泡巨噬细胞和原代肺细胞中的UDS。一项时间进程研究的数据显示,在用[3H]胸腺嘧啶孵育16小时后,实现了高放射性标记的DNA修复。在低血清(1%)的条件下,浓度为20 mM的羟基脲以令人满意的方式抑制了原代肺细胞中的正常DNA合成(抑制率为81 - 88%)。按照此方案,N - 甲基 - N'- 硝基 - N - 亚硝基胍和2 - 氨基蒽在大鼠肺泡巨噬细胞和原代肺细胞中均诱导了与剂量相关的UDS增加。结果表明,培养的原代大鼠肺细胞具有DNA修复能力,并且UDS试验可能有助于评估该细胞系统中化学物质的肺基因毒性作用。

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