香烟烟雾对A549与A549-R细胞的氧化损伤

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目的探讨hOGG1基因低表达对香烟烟雾作用下细胞氧化损伤效应的影响。方法不同浓度香烟烟雾暴露A549细胞和hOGG1基因低表达的A549-R细胞,MTT实验检测两种细胞存活率,彗星实验观察两种细胞DNA损伤,荧光法测定两种细胞内活性氧(ROS)含量,高效液相色谱-电化学法(HPLC-ECD)检测细胞基因组DNA中8-羟基脱氧鸟苷(8-OHdG)的含量。结果随着香烟烟雾暴露浓度的增加,两种细胞的存活率均下降,且A549-R细胞IC50显著小于A549细胞,差异有统计学意义(P<0.05);两种细胞ROS生成量与香烟烟雾浓度呈剂量-效应关系,当香烟烟雾浓度≥1.25支/升时,A549-R细胞ROS含量显著高于A549细胞(P<0.05);不同香烟烟雾浓度下,A549-R细胞拖尾率、尾长、OTM值均大于A549细胞,差异有统计学意义(P<0.05);A549细胞与A549-R细胞基因组DNA8-OHdG含量随香烟烟雾浓度的增加而升高,当香烟烟雾浓度为2.5支/升和5支/升时,A549-R细胞8-OHdG含量显著高于A549细胞(P<0.05)。结论香烟烟雾可导致A549细胞与A549-R细胞的氧化损伤,hOGG1基因低表达可增加A549-R细胞对香烟烟雾引起的氧化损伤的敏感性。 Objective To investigate the effect of low expression of hOGG1 gene on cell oxidative damage induced by cigarette smoke. Methods A549 cells exposed to different concentrations of cigarette smoke and low expression of hOGG1 gene in A549-R cells were detected by MTT assay, DNA damage in both cells was detected by comet assay, and the content of reactive oxygen species (ROS) , 8-OHdG (8-OHdG) in cell genomic DNA was detected by HPLC-ECD. Results With the increase of cigarette smoke exposure, the survival rate of both cells decreased, and the IC50 of A549-R cells was significantly lower than that of A549 cells (P <0.05). The ROS production of both cells was significantly lower than that of cigarette smoke The concentration of ROS in A549-R cells was significantly higher than that of A549 cells (P <0.05) when the concentration of cigarette smoke was more than 1.25, and the tailing rate of A549-R cells (P <0.05). The content of 8-OHdG in A549 cells and A549-R cells increased with the increase of cigarette smoke concentration. When the cigarette smoke concentration was 2.5 / The content of 8-OHdG in A549-R cells was significantly higher than that in A549 cells (P <0.05). Conclusion Cigarette smoke can induce oxidative damage of A549 cells and A549-R cells. Low expression of hOGG1 gene can increase the sensitivity of A549-R cells to cigarette smoke-induced oxidative damage.
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