香烟烟雾对人支气管上皮细胞的氧化损伤研究  

Cigarette Smoke-induced Oxidative Damage to Human Bronchial Epithelial Cells

作　　者：刘蜀坤[1] 王玲[1] 陈晨[1] 张遵真[1] 

机构地区：[1]四川大学华西公共卫生学院环境卫生教研室,成都610041

出　　处：《四川大学学报（医学版）》2009年第4期667-671,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30571535和30872079)资助

摘　　要：目的探讨香烟烟雾对人支气管上皮细胞氧化损伤的影响,评价氧化损伤效应在香烟烟雾致肺癌中的作用。方法体外培养人支气管上皮细胞(HBE),以DMSO作为吸收液采集香烟主流烟雾,MTT比色法测量香烟烟雾对HBE细胞的毒性,彗星实验和微核实验分别检测细胞DNA链断裂和细胞染色体损伤,同时使用荧光探针测定细胞内活性氧(ROS)的含量。结果随着香烟烟雾浓度的增加,HBE细胞存活率逐渐下降;随着染毒时间的延长,香烟烟雾对HBE细胞的半数抑制浓度(IC50)逐渐降低,显示明显的剂量-效应和时间-效应关系。香烟烟雾可以诱导HBE细胞DNA链断裂,表现为细胞拖尾率增加,同时尾长、尾部DNA含量和Olive尾距均随香烟烟雾浓度增加而增大。香烟烟雾亦可诱导HBE细胞染色体损伤,当香烟烟雾浓度≥1支/L时,实验组微核率与对照组比差异均有统计学意义(P<0.05)。此外,HBE细胞ROS生成量与香烟烟雾浓度呈剂量-效应关系。结论香烟烟雾可以直接诱导人支气管上皮细胞ROS增加,从而造成细胞毒性增加、染色体损伤和DNA链断裂,提示氧化损伤是香烟烟雾致肺癌的重要机制。Objective To determine the oxidative damage of mainstream smoke （MS） on Human bronchial epithelial cells （HBE） and its role in lung cancer. Methods MTT assay was used to test the cytotoxicity of MS on HBE. The HBE cells were treated with different concentrations of MS for 12 h. The chromatosome damage and DNA strand breaks were measured by micronucleus test and alkaline comet assay respectively. The contents of ROS in the HBE cells were determined using fluorescence method. Results With the increase of MS, the viability of HBE cells decreased. The IC50 decreased with the increasing exposure time to MS, showing significant dose-effect and time-effect relationships. The MS induced DNA strand break in the HBE cells. The comet, L Tail, Tail DNA and OTM increased with the increase of MS concentrations. Cigarette smoke also induced chromosome damage. The micronucleus rate of the HBE ceils exposed to more than one cigarette/L of MS was significantly greater than the controls （P〈0.05）. The ROS increased with the concentration of MS. Conclusion MS induces ROS in HBE cells, resulting in increased cytotoxicity, chromosome damage and DNA strand breaks, which suggests that oxidative damage is an important mechanism of lung cancer caused by MS.

关 键 词：主流烟雾 人支气管上皮细胞 活性氧 DNA损伤 

分 类 号：R114[医药卫生—卫生毒理学]