亚砷酸As(Ⅲ)诱导人肺癌A549细胞氧化应激反应的实验研究  

Induction of oxidative stress by arsenious acid As(Ⅲ)in A549 cells

作　　者：赵俊伟[1] 华辰凤 尚平平[1] 谢复炜[1] 李翔[1] ZHAO Junwei;HUA Chenfeng;SHANG Pingping;XIE Fuwei;LI Xiang(Key Laboratory of Tobacco Chemistry,Zhengzhou Tobacco Research Institute of China National Tobacco Corporation,Zhengzhou 450001,Henan,China)

机构地区：[1]中国烟草总公司郑州烟草研究院烟草化学重点实验室,河南郑州450001

出　　处：《癌变．畸变．突变》2021年第1期28-31,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：郑州烟草研究院院长科技发展基金(322014CA0350)。

摘　　要：目的:探讨亚砷酸As(Ⅲ)对人肺癌A549细胞氧化应激的诱导作用。方法:将处于对数生长期的A549细胞暴露于不同浓度的亚砷酸As(Ⅲ)溶液(以砷浓度计分别为0、2.5、5、10和20μg/mL)中培养24 h,采用CCK-8法检测细胞存活率;用2.5和5μg/mL的亚砷酸As(Ⅲ)溶液染毒A549细胞24 h,并设对照组(含10%胎牛血清的RPMI-1640培养基),采用ELISA法检测细胞培养液中的超氧化物歧化酶(SOD)浓度,荧光探针DCFH-DA检测细胞内活性氧(ROS)的表达水平。结果:随着亚砷酸As(Ⅲ)溶液染毒浓度的升高,A549细胞的存活率呈逐渐降低的趋势(r=0.99,P<0.05)。2.5和5μg/mL亚砷酸As(Ⅲ)溶液染毒24 h后,细胞培养液中SOD的浓度分别为(0.72±0.05)、(1.10±0.16)ng/mL,均高于对照组的(0.56±0.03)ng/mL(P<0.05);细胞内ROS的相对荧光强度分别为(169.31±6.13)%和(242.60±2.35)%,均高于对照组的(100±2.80)%(P<0.05)。结论:A549细胞暴露于亚砷酸As(Ⅲ)溶液可致细胞存活率降低,细胞培养液中SOD和细胞内ROS浓度均升高;氧化应激可能是亚砷酸As(Ⅲ)致A549细胞毒性的机制之一。OBJECTIVE:To investigate the induction of oxidative stress by arsenious acid solution As(Ⅲ) in human lung cancer A549 cells.METHODS:Cultured A549 cells were exposed to arsenious acid solution,As(Ⅲ),at doses of 0,2.5,5,10 and 200 μg/mL for 24 h.Cell viabilities were determined using the CCK-8 assay.For cells exposed to 0,2.5,5 μg/mL doses for 24 h,superoxide dismutase(superoxide dismutase,SOD) in cell culture medium was detected using ELISA,and intracellular reactive oxygen species(ROS) were detected using fluorescent probe DCFH-DA.RESULTS:As exposure concentration of As(Ⅲ)increased,A549 cell viabilities were gradually decreased(r=0.99,P<0.05).The level of SOD in the 2.5,and 5 μg/mL exposure groups were(0.72±0.05),and(1.10±0.16) ng/mL,respectively,which were higher than that of the control group [(0.56±0.03) ng/mL](P<0.05).Relative fluorescence intensities of ROS were(169.31±6.13)%,and(242.60±2.35)% for the same two groups which were higher than that of the control group(100±2.80)%.CONCLUSION:As(Ⅲ) exposure caused significantly reduced cell survival rates,increased concentrations of SOD and ROS in A549 cells.Oxidative stress may therefore be an important mechanism for the toxicity of As(Ⅲ).

关 键 词：亚砷酸 氧化应激 人肺癌A549细胞 超氧化物歧化酶 活性氧 

分 类 号：R114[医药卫生—卫生毒理学]