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N-乙酰半胱氨酸对辐射致小鼠骨髓单个核细胞损伤的保护作用

张俊伶1,路璐2,李德冠2,吴红英2,孟爱民2   

  1. 1. 中国医学科学院放射医学研究所
    2. 天津市医学科学院放射医学研究所
  • 收稿日期:2013-06-03 修回日期:2013-09-04 出版日期:2014-01-15 发布日期:2014-01-15
  • 通讯作者: 孟爱民

Protective effect of N-acetyl-cysteine on irradiation-induced bone marrow mononuclear cells injury

  • Received:2013-06-03 Revised:2013-09-04 Published:2014-01-15 Online:2014-01-15

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摘要： 目的 观察抗氧化剂N-乙酰半胱氨酸 （NAC） 对辐射致小鼠骨髓单个核细胞 （BMMNCs） 损伤的保护作用， 进一步探讨其可能的作用机制。方法 实验分为对照组、 辐射组 （辐射剂量1、 4 Gy）、 辐射加NAC组 （辐射1、 4 Gy前30 min各组加入NAC）。按分组不同将2×106/mL浓度细胞悬液与RPMI-1640培养基和2×10-5mol/L NAC溶液加入2 mL无菌EP管内， 37 ℃ CO2孵箱内孵育30 min后γ射线分别辐射1、 4 Gy。生物发光法检测小鼠BMMNCs细胞活力、活性氧检测探针 （DCFH-DA） 检测细胞内活性氧 （ROS） 水平、 集落形成数目检测克隆形成能力。结果 辐射暴露4Gy后， 辐射组小鼠BMMNCs细胞活力 （发光值： 284 296.7±16 541.2）， 明显低于对照组的 （848 586.7±61 404.4）； 辐射暴露1 Gy后， 辐射组细胞内ROS水平 （荧光值： 6 750.0±103.5） 高于对照组的 （5 710.7±56.2）， 小鼠BMMNCs克隆形成能力辐射组为 （626.7±51.3） 克隆数/105细胞， 低于对照组的 （986.7±100.7） 克隆数/105细胞； 而辐射加NAC组， 与辐射组比较小鼠BMMNCs细胞活力增加至发光值352 770.0±23 466.1、 细胞内ROS水平下降至荧光值5430.0±61.0、BMMNCs克隆形成能力增加至 （773.3±49.3） 克隆数/105细胞。结论 NAC对辐射暴露小鼠BMMNCs损伤可能是通过降低细胞内ROS水平达到保护作用。NAC可以作为后续实验研究的阳性对照药。

关键词: 乙酰半胱氨酸, 单核细胞, 骨髓, 辐射损伤, 辐射剂量, 辐射防护, 小鼠, 近交C57BL/6

Abstract: Objective To observe N-acetyl-cysteine's protective effect on irradiation-induced bone marrow mononuclear cells(BMMNCs) injury, and explore the possible mechanism. Methods There were 3 groups in the research: control group,irradiation group(doses of irradiation were 1Gy and 4Gy) and irradiation with NAC group(The NAC was cocultured with BMMNCs half hour before irradiation).We added 2×106/mL BMMNCs and the RPMI-1640 medium or 2×10-5 mol/L NAC into the 2mL EP tubes according to the different requirement of groups,then all the tubes were cultured in 37 ℃ CO2 incubator for 30 mins and irradiated 1Gy and 4Gy. Mouse BMMNCs 'cell viability was measured by bioluminescence, the level of cell reactive oxygen species was measured by DCFH-DA, and the ability of colony-forming units was reflected by CFU-GM. Results After 4Gy irradiation exposure,the BMMNCs 'cell viability was (284 296.7±16 541.2) Relative Light Units,which was lower than the control group(848 586.7±61 404.4) Relative Light Units. And After 1Gy irradiation exposure , the level of cell reactive oxygen species was （6 750.0±103.5）, which was higher than control group(5 710.7±56.2)Fluorescence Value; The number of colony-forming units per 105 cells after irradiation exposure was (626.7±51.3),significantly lower than the control group (986.7±100.7).Compared to the irradiation group, when administrate NAC half hour before irradiation, the BMMNCs 'cell viability increased to (352 770.0±23 466.1) Relative Light Units , the level of cell reactive oxygen species decreased to (5 430.0±61.0) Fluorescence Value , the number of colony-forming units per 105 cells increased to(773.3±49.3),and the result has significantly difference. Conclusion NAC has protective effect on irradiation-induced BMMNCs injury，it can play the role of positive control for the following work.

Key words: acetylcysteine, monocytes, bone marrow, radiation injuries, radiation dosage, radiation protection, mice, 近交C57BL/6