不同粒径谱汽油发动机尾气气-液界面染毒对BEAS-2B细胞存活率的影响

Evaluation of the viability of BEAS-2B cells exposed to gasoline engine exhaust with different particle sizes by air-liquid interface

目的 采用气-液界面（ALI）体外细胞染毒技术,评价不同粒径谱汽油发动机尾气（GEE）对人肺BEAS-2B细胞存活率的影响.方法 用Tedlar集气袋收集GEE,并通过TSI-3321和SMPS-3938两种粒径谱仪分析过滤后的汽车尾气组、汽车尾气组和非三元催化配置的摩托车尾气组的颗粒物个数、表面积和质量浓度,同时设置空白对照组和洁净空气组.除了空白对照组,其余4组采用ALI体外细胞染毒技术,在染毒流量为25 ml/min,37℃水浴条件下,对生长在Transwell插件多孔膜上的BEAS-2B细胞持续染毒60 min.通过CCK-8细胞毒性检测试剂盒测定细胞的相对存活率.结果 过滤后的汽车尾气组、汽车尾气组和摩托车尾气组均可检出较高浓度的细颗粒物,但粗颗粒物很少,且呈摩托车尾气组〉汽车尾气组〉过滤后的汽车尾气组的趋势（P〈0.001）.与洁净空气组比较,过滤后的汽车尾气组、汽车尾气组和摩托车尾气组的细胞相对存活率均明显降低,差异有统计学意义（P〈0.001）.各GEE暴露组之间的比较显示,细胞相对存活率呈过滤后的汽车尾气组〉汽车尾气组〉摩托车尾气组的趋势（P〈0.001）;以洁净空气组为参考,过滤后的汽车尾气组、 汽车尾气组、 非三元催化配置的摩托车尾气组的细胞相对存活率均数依次降低了26.34%、36.00%和49.59%.结论 人肺BEAS-2B细胞ALI暴露于不同粒径谱的GEE可产生不同程度的毒性作用,降低GEE颗粒物浓度以及配置三元催化装置可明显改善肺细胞的存活率.

Objective To evaluate the viability of gasoline engine exhaust （GEE） with different particle sizes on human lung cell line BEAS-2B in vitro by air-liquid interface （ALI）. Methods GEE were collected with a Tedlar bag and their particulate matter （PM） number, surface and mass concentration in three kind of GEE （filtered automobile exhaust, non-filtered automobile exhaust and motorcycle exhaust without three-way catalytic converter） were measured by two type of particle size spectrometer including TSI-3321 and SMPS-3938. Five groups were included, which divided into blank control group, clean air group, filtered automobile exhaust group, non-filtered automobile exhaust group and motorcycle exhaust without three-way catalytic converter group. Except the blank control group, BEAS-2B cells, cultured on the surface of Transwells, were treated with clean air or GEE by ALI method at a flow rate of 25 ml/min, 37℃for 60 min in vitro. CCK-8 cytotoxicity test kit was used to determine the cell relative viability of BEAS-2B cells. Results In the filtered automobile exhaust, non-filtered automobile exhaust and motorcycle exhaust without three-way catalytic converter , high concentrations of fine particles can be detected, but the coarse particles only accounted for a small proportion, and the sequence of PM concentration was motorcycle exhaust without three-way catalytic converter group〉 non-filtered automobile exhaust group〉filtered automobile exhaust group （P〈0.001）. Compared with the clean air group, the cell relative viability in the 3 GEE-exposed groups were significantly lower （P〈0.001）. Among the comparisons of GEE exposure groups with different particle size spectra , the sequence of the cell relative viability was filtered automobile exhaust group 〉non-filtered automobile exhaust group〉motorcycle exhaust without three-way catalytic converter group （P〈0.001）. When took the clean air control group as a reference, the mean of the cell relative viability in the filtered automobile exhaust group , non-filtered automobile exhaust group and motorcycle exhaust without three-way catalytic converter group , was decreased by 26.34%, 36.00% and 49.59%, respectively. Conclusion GEE with different particle size spectra could induce different levels of toxic effects to the human lung cells BEAS-2B by ALI. After lowering the concentration of particles in the GEE and using the three-way catalytic converter could obviously improve the survival rate of lung cells.