摩托车尾气气-液界面暴露对人体呼吸道不同细胞存活率和细胞器的影响  

Effects of motorcycle exhaust on cell relative viabilities and organelles damage in human respiratory tract by air-liquid interface exposure

作　　者：周玉梅 鱼涛[1] 张雪艳[1] 刘黎[1] 李斌[1] 戴宇飞[1] 宾萍[1] Zhou Yumei;Yu Tao;Zhang Xueyan;Liu Li;Li Bin;Dai Yufei;Bin Ping(National Institute of Occupational Health and Poison Control,Chinese Center for Disease Control and Prevention,Beijing 100050,China)

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,北京100050

出　　处：《卫生研究》2022年第2期271-277,共7页Journal of Hygiene Research

基　　金：国家自然科学基金面上项目(No.82073596,81472955)。

摘　　要：目的应用气-液界面(air-liquid interface,ALI)染毒技术,探讨摩托车尾气(motorcycle exhaust,ME)对呼吸道不同细胞的细胞器损伤和相对存活率的影响。方法用20 L Tedlar气袋收集洁净空气和ME。实验分为空白对照组、洁净空气组和ME组。空白对照组未做任何处理,洁净空气组为连接0.2μm滤器后收集的含有21%O_(2)和79%N_(2)的人造空气,ME组为用气袋直接收集的ME。除了空白对照组,其余两组利用ALI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ALI插件多孔膜上的BEAS-2B、A549和人肺巨噬细胞持续染毒60 min。利用透射电镜观察细胞内细胞器的损伤。利用CCK-8(cell counting kit-8)检测法评价细胞的相对存活率。结果细胞电镜显示,对照组和洁净空气组的三种细胞细胞膜均为完整的单层膜结构;线粒体双层膜完整,线粒体嵴清晰可见;核内染色质分布均匀;洁净空气组中偶见少量自噬小体。ME暴露组中,可见BEAS-2B、A549和人肺巨噬细胞的细胞核膜萎缩,线粒体肿胀并伴有线粒体嵴消解和断裂现象,自噬小体的数量明显增多,自噬小体内可见吞噬的细胞器和众多膜泡状结构。BEAS-2B、A549和人肺巨噬细胞相对存活率分别为(61.65±6.23)%、(46.55±6.12)%和(40.89±1.59)%,与洁净空气组相比,3种细胞存活率均显著下降(P<0.01)。方差分析提示,3种细胞存活率差异有统计学意义(F=62.91,P<0.01),细胞相对存活率呈现BEAS-2B>A549>人肺巨噬细胞的趋势。结论BEAS-2B、A549和人肺巨噬细胞在气-液界面暴露于ME后可引起呼吸道细胞器结构的改变、细胞相对存活率的下降,对呼吸道不同细胞的细胞器和相对存活率存在显著影响。OBJECTIVE Applying air-liquid interface(ALI)technology,learn the effect of motorcycle exhaust(ME)on relative viabilities and organelle damages of different cells in the respiratory tract.METHODS Use 20 L Tedlar airbags to collect clean air and ME.Three groups were included,which divided into blank control group,clean air group and ME group.The blank control group did not do any treatment,the clean air group was used artificial air containing 21%O_(2) and 79%N_(2) and collect after connecting with a 0.2μm particle filter,the ME group was used ME directly collected with air bags.Except the blank control group,BEAS-2 B,A549 and human lung macrophages which grown on the membrane were treated with clean air or ME by ALI exposure at a flow rate of 25 mL/min,37℃for 60 min in vitro.Use transmission electron microscope(TEM)to observe the damage of intracellular organelles.Cell relative viability was evaluated by cell counting kit-8 assay.RESULTS As the cytoelectron microscopy showed,in blank control group and clean air group,the mitochondrial bilayer membranes of the three types of cells were intact;the mitochondrial cristae was clearly visible and the chromatin in the nucleus was evenly distributed.But a few autophagosomes were occasionally found in the clean air group.In the ME group,the nuclear membranes of BEAS-2 B,A549 and human lung macrophages shrunk,mitochondria were swollen with mitochondrial cristae degraded and broken,autophagosomes increased significantly meanwhile observed the organelles and many vesicle like structures in autophagosomes.The relative viabilities of BEAS-2 B,A549 and human lung macrophages were(61.65±6.23)%,(46.55±6.12)%and(40.89±1.59)%,respectively.Compared with the clean air group,all of the relative vibilities in the three kinds of cells were decreased significantly(P<0.01).The analysis of variance showed that there was a significant difference in the relative viabilities of the three kinds of cells(F=62.91,P<0.01),and the relative viabilities of the cells showed a trend of BEAS-2 B>A

关 键 词：摩托车尾气 气-液界面 呼吸道细胞 细胞电镜 

分 类 号：X51[环境科学与工程—环境工程] X503.1