杭州市PM2.5对BEAS-2B细胞的生物学效应研究  

Assessment of biological effects in BEAS-2B cells exposed to PM2.5 collected in Hangzhou City

作　　者：吕烨[1] 徐虹[1] 徐珊珊[1] 朱冰[1] 高兵 章美 LV Ye;XU Hong;XU Shan-shan;ZHU Bing;GAO Bing;ZHANG Mei(Hangzhou Municipal Center for Disease Control and Prevention,Hangzhou,Zhejiang 310021,China;不详)

机构地区：[1]杭州市疾病预防控制中心,浙江杭州310021 [2]淳安县疾病预防控制中心,浙江淳安311700

出　　处：《中国卫生检验杂志》2022年第14期1691-1694,1734,共5页Chinese Journal of Health Laboratory Technology

摘　　要：目的观察杭州市不同地区PM2.5对人支气管上皮细胞(BEAS-2B)的生物学效应。方法采集杭州市下城区和淳安县的PM2.5样品,通过超声冻干提取后将不同浓度PM2.5样品作用于BEAS-2B细胞,24 h后观察其生物学效应。用CCK-8法检测细胞活力,根据实验结果,将细胞分成空白对照组、25μg/ml浓度组和100μg/ml浓度组。采用γH2AX免疫荧光法和免疫印迹法分别检测γH2AX焦点损伤及γH2AX蛋白表达情况,免疫荧光实验采用平均焦点数和焦点阳性细胞率作为DNA损伤的评价指标;用DCFH-DA探针法观察细胞内ROS生成。结果CCK-8实验结果显示,下城区和淳安县的PM2.5样品在25μg/ml、50μg/ml、100μg/ml、200μg/ml和400μg/ml浓度时能显著降低细胞活力(P<0.05),而2.5μg/ml~10μg/ml低浓度作用则未对细胞造成影响。用25μg/ml和100μg/ml浓度PM2.5样品处理细胞后,细胞内γH2AX平均焦点数、焦点阳性细胞率、γH2AX蛋白以及ROS表达均有显著增加(P<0.05)。结论PM2.5能使BEAS-2B细胞活力降低、细胞DNA损伤,具有一定的细胞毒性,并能使细胞内ROS表达增强。Objective To assess the biological effects of PM2.5 on human bronchial epithelial cells(BEAS-2B).Methods PM2.5 were collected from Xiacheng District and Chun’an County,and extracted by ultrasonic freeze-drying.ThenBEAS-2B cells were exposed to different concentrations of PM2.5.After a CCK-8 test,the experiments were divided intothree groups:sham group,25μg/ml group and 100μg/ml group.After exposure,cells were subjected toγH2AX immunofluo-rescence and western blot.In the immunofluorescence experiment,the average number of foci per cell and the percentage offoci-positive cells were used as the indicators of DNA damage.Reactive oxygen species(ROS)production was assessed by theDCFH-DA probe.Results The results of CCK-8 showed that PM2.5 concentration in Xiacheng District and Chun’an Countycould significantly reduce cell viability at 25μg/ml,50μg/ml,100μg/ml,200μg/ml and 400μg/ml(P<0.05).However,low concentrations of 2.5μg/ml-10μg/ml did not affect the cells.After treated with 25μg/ml and 100μg/mlPM2.5 samples,the average number of focal points ofγH2AX,the rate of focal positive cells,the expression ofγH2AX proteinand ROS were significantly increased(P<0.05).Conclusion PM2.5 can reduce the activity of BEAS-2B cells,damagecell DNA,express certain cytotoxicity,and enhance intracellular ROS expression.

关 键 词：PM2.5 DNA损伤 γH2AX 活性氧自由基 

分 类 号：R73[医药卫生—肿瘤]