气-液界面暴露汽车尾气致BEAS-2B细胞的毒性效应
发布时间：2018-01-15 12:29
本文关键词：气-液界面暴露汽车尾气致BEAS-2B细胞的毒性效应 出处：《卫生研究》2017年05期 论文类型：期刊论文
【摘要】：目的应用新型的气-液界面细胞染毒手段,探讨汽车尾气对人肺细胞毒性效应的影响程度,以及采用0.2μm滤器过滤尾气干预后的细胞毒性变化情况。方法用20 L Tedlar气袋收集汽车尾气,并通过粒径谱仪分析0.2μm滤器过滤前后的汽车尾气颗粒物数量、表面积和质量浓度。实验分为空白对照组、洁净空气组、汽车尾气过滤组和汽车尾气组。空白对照组未作任何处理;洁净空气组为21%O_2和79%N_2;汽车尾气过滤组为用0.2μm颗粒物过滤器过滤后的汽车尾气;汽车尾气组为用气袋直接收集的汽车尾气。除了空白对照组,其余3组利用气-液界面体外染毒技术,在染毒流量为25 mL/min,37℃水浴条件下,对生长在气-液界面插件多孔膜上的BEAS-2B细胞持续染毒60 min。利用CCK-8(cell counting Kit-8)检测法评价细胞的存活率。采用2',7'-二氯荧光黄双乙酸盐(2',7'-dichloro-dihydro-fluorescein diacetate,DCFH-DA)探针检测细胞内活性氧(reactive oxygen species,ROS)水平。Annexin V-FITC/PI细胞凋亡检测试剂盒检测汽车尾气对BEAS-2B细胞凋亡及坏死率的影响。结果汽车尾气组中0.5~10μm颗粒物数量浓度、表面积浓度和质量浓度分别为0.24×10~3个/cm~3、0.29×10~3μm~2/cm~3和0.19μg/m~3,10~500 nm颗粒物数量浓度、表面积浓度和质量浓度分别为56×10~3个/cm~3、34.53×10~8nm~2/cm~3和95 ng/m~3。汽车尾气过滤组0.5~10μm颗粒物数量浓度、表面积浓度和质量浓度分别低于1个/cm~3、1μm~2/cm~3和0.001μg/m~3,10~500 nm颗粒物数量浓度、表面积浓度和质量浓度与汽车尾气组相比分别降低了89.79%、93.57%和90.55%。洁净空气组的细胞存活率、胞内ROS、早期凋亡率和晚期凋亡坏死率分别为(90.15±4.25)%、(1.92±0.34)×10~5、(1.09±0.48)%、(8.93±3.31)%。与洁净空气组比较,两组汽车尾气组的细胞存活率、胞内ROS和晚期凋亡及坏死率差异均有统计学意义(P0.05)。汽车尾气过滤组与汽车尾气组之间的比较发现,汽车尾气组细胞存活率[(58.09±4.06)%]低于汽车尾气过滤组[(66.85±3.49)%],差异有统计学意义(t=6.331,P0.001),胞内ROS[汽车尾气过滤组∶汽车尾气组=(2.94±0.21)×10~5∶(3.32±0.49)×10~5,t=-1.252,P=0.279]、早期凋亡率[汽车尾气过滤组∶汽车尾气组=(1.09±0.30)%∶(0.99±0.10)%,t=0.708,P=0.497]和晚期凋亡坏死率[汽车尾气过滤组∶汽车尾气组=(21.75±10.37)%∶(15.32±2.74)%,t=1.347,P=0.242]差异均无统计学意义(P0.05)。结论人肺细胞气-液界面暴露于汽车尾气可产生显著的毒性效应,0.2μm过滤器过滤尾气干预后,细胞毒性会明显降低。
[Abstract]:Objective to investigate the effect of automobile exhaust on the cytotoxicity of human lung cells by means of a new method of cell exposure at the gas-liquid interface. And the changes of cytotoxicity after the treatment with 0.2 渭 m filter filter. Methods the vehicle exhaust was collected by 20L Tedlar air bag. The quantity, surface area and mass concentration of vehicle exhaust particles before and after filtration with 0.2 渭 m filter were analyzed by particle size spectrometer. The experiment was divided into blank control group and clean air group. Vehicle exhaust filtration group and vehicle exhaust group. Blank control group did not do any treatment; In the clean air group, there were 21 O / s _ 2 and 79 / N _ 2; The vehicle exhaust was filtered by 0.2 渭 m particulate matter filter. The vehicle exhaust group was collected directly by air bag. In addition to the blank control group, the other three groups were exposed to the gas liquid interface in vitro under the water bath condition of 25 mL 路min ~ (-1) 路min ~ (-1) ~ 37 鈩，
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