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  

Protective Effects of N-Acetyl-Cysteine on Irradiation-Induced Bone Marrow Mononuclear Cell Injury

作　　者：张俊伶[1] 路璐[1] 李德冠[1] 吴红英[1] 孟爱民[1] 

机构地区：[1]中国医学科学院放射医学研究所,天津市放射医学与核医学重点实验室,300192

出　　处：《天津医药》2014年第1期54-57,共4页Tianjin Medical Journal

基　　金：国家973重大专项资助项目(项目编号:2011CB964800-G);国家自然科学基金资助项目(项目编号:81072237,81129020);天津市重点基金资助项目(项目编号:11JCZDJC19100);教育部高等学校博士学科点专项基金(项目编号:20111106110038);中国医学科学院放射医学研究所发展基金(项目编号:1309)

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5 430.0±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。Objective To observe the protective effect of N-acetyl-cysteine （NAC） on the injury of irradiation-in- duced bone marrow mononuclear cells （BMMNCs）, and explore the possible mechanism. Methods There were 3 groups in the study： control group, irradiation group （doses of irradiation were 1 Gy and 4 Gy） and irradiation with NAC group （NAC was cocuhured with BMMNCs half hour before irradiation）. The 2xlO6/mL BMMNCs and the RPMI-1640 medium or 2x10-5 mol/L NAC were added into the 2 mL EP tubes according to the different requirement of groups. The tubes were then cul- tured in the 37 ~C CO2 incubator for 30 min and irradiated with 1 Gy and 4 Gy. The viability of BMMNCs was measured by bioluminescence. The level of reactive oxygen species （ROS） was measured by DCFH-DA, and the ability of colony-forming units was detected by CFU-GM. Results After 4 Gy irradiation exposure, the cell viability of BMMNCs was significantly lower in irradiation group （284 296.7＋16 541.2） than that of control group （848 586.7＋61 404.4）. After 1 Gy irradiation expo- sure, the level of ROS was higher in irradiation group （6 750.0＋ 103.5） than that of control group （5 710.7＋56.2）. The number of colony-forming units per 105 cells after irradiation exposure was （626.7＋51.3）, which was significantly lower than that of control group （986.7＋ 100.7）. Compared to irradiation group, the cell viability of BMMNCs increased to （352 770.0＋23 466.1） in irradiation with NAC group. The level of ROS decreased to （5 430.0＋61.0）, and the number of colony-forming units per 105 cells increased to （773.3±49.3）. Conclusion NAC has protective effect on irradiation-induced injury in BMMNCs, which may be related with the decreased level of ROS. NAC can play the role of positive control for the following work.

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 

分 类 号：R551[医药卫生—血液循环系统疾病]