摘要
目的比较4种大气颗粒物对人Ⅱ型肺泡上皮细胞A549的毒性效应。方法将收集的新乡市冬季大气细颗粒物(PM 2.5)与常用的标准参考颗粒物(柴油机尾气颗粒物DEP-Sagai、SRM 2975及城区扬尘标准颗粒物SRM 1649)分别用磷酸盐缓冲液制备成颗粒物悬液,对人肺泡上皮细胞株A549进行24 h不同剂量染毒,采用甲基噻唑基四唑比色法、乳酸脱氢酶(LDH)测定试剂盒、活性氧(ROS)检测试剂盒及人白细胞介素-8(IL-8)酶联免疫吸附试验试剂盒分别检测细胞相对存活率、细胞培养上清液中LDH和IL-8水平、细胞内ROS水平。结果4种颗粒物均可致A549细胞损伤,且该效应呈剂量依赖性。低剂量SRM 1649即可明显引起细胞死亡,当剂量超过30 mg·L^-1后其细胞存活率趋于稳定。4种颗粒物处理的细胞培养上清液中的LDH水平自高至低依次为SRM 2975>SRM 1649>DEP-Sagai>PM 2.5,IL-8水平自高至低依次为PM 2.5>DEP-Sagai>SRM 2975>SRM 1649;4种颗粒物均可不同程度地诱导A549细胞内ROS水平升高,在相同剂量下4种颗粒物诱导ROS产生的能力依次为DEP-Sagai>SRM 2975>SRM 1649>PM 2.5。结论4种大气颗粒物在受试剂量范围内均能不同程度地引起A549细胞损伤、氧化应激和炎症反应,且呈剂量效应。
Objective To study the toxicological effects of different types of ambient particulate matter on human alveolar epithelial cell line A549.Methods The suspension of PM 2.5 which were collected during the winter of Xinxiang city,diesel exhaust particles including DEP-Sagai and SRM 2975,and urban dust SRM1649 were prepared by phosphate buffer solution,and then the A549 cells line were stimulated by different doses of suspension of four particulate matter for 24 hours.The relative survival rate of cells,the levels of lactate dehydrogenase(LDH),interleukin-8(IL-8)in cell culture supernatant and reactive oxygen species(ROS)level in cells were detected by methl thiazolyl tetrazolizum colorimetry,LDH test kit,ROS test kit and human enzyme-linked immunosorbent kit respectively.Results All four kinds of particulate matter could cause damage to A549 cells,and the effect was dose dependent.SRM 1649 could caused cell death at low dose,and its cell survival rate stabilized when the dose exceeded 30 mg·L^-1.The LDH level in the cell culture supernatants of the four particulates was SRM 2975>SRM 1649>DEP-Sagai>PM 2.5,and the IL-8 level was PM 2.5>DEP-Sagai>SRM 2975>SRM 1649;four kinds of particulate matter could induce the increase of ROS level in A549 cells in different extents,and the induction potencies of four kinds of particulate matter at the same dose was DEP-Sagai>SRM 2975>SRM 1649>PM 2.5.Conclusion Four kinds of particulate matter can cause damage to A549 cells within the range of reagent dosage,and it has a dose effect.
作者
朱静芳
田歌
范威
刘影影
张艳格
王贵
吴卫东
ZHU Jing-fang;TIAN Ge;FAN Wei;LIU Ying-ying;ZHANG Yan-ge;WANG Gui;WU Wei-dong(College of Public Health,Xinxiang Medical University,International Joint Laboratory on Health Effects and Intervention of Air Pollution,,Xinxiang 453003,Henan Province,China)
出处
《新乡医学院学报》
CAS
2019年第11期1007-1012,共6页
Journal of Xinxiang Medical University
基金
国家自然科学基金资助项目(编号:81573112)
国家重点研发计划项目(编号:2016YFC0900803,2017YFD0400301)
关键词
大气颗粒物
肺泡上皮细胞
毒性效应
活性氧
白细胞介素-8
particulate matter
alveolar epithelial cells
poisoning effect
reactive oxygen species
interleukin-8
