利用实时细胞分析法和Alamar Blue法测定雾霾细颗粒物对呼吸系统的细胞毒性  

Study on cytotoxicity of respiratory system exposed to fine particulate matter by real time cell analyze and Alamar Blue

作　　者：顾雯[1] 张少平[1] 李毅民[1] 段链[1] 

机构地区：[1]中国疾病预防控制中心环境与健康相关产品安全所,北京100021

出　　处：《卫生研究》2016年第5期829-833,共5页Journal of Hygiene Research

基　　金：卫生行业专项(No.201402022)

摘　　要：目的利用实时细胞分析(real time cell analyze,RTCA)法及Alamar Blue法测定雾霾细颗粒物对于人支气管上皮细胞(BEAS-2B)和人胚肺成纤维细胞(MRC-5)的毒性作用,比较两种方法检测雾霾细颗粒物对呼吸系统影响的差异。方法分别将BEAS-2B及MRC-5细胞接种于RTCA配套16孔板及普通96孔板。接种24 h后,分别用终浓度为2500、1250、625、312.5、156.25和78.125μg/m L的雾霾细颗粒物进行染毒。选择3、12、24、36和48 h通过RTCA法及Alamar Blue法两种方法进行测定,计算IC_(50)。采用SPSS 16.0,利用配对t检验,对两种方法所得IC_(50)进行统计分析,并分析其相关性。结果雾霾细颗粒物作用于两种细胞后分别利用RTCA法及Alamar Blue法测得的IC50具有相关性,但差异无统计学意义。结论雾霾细颗粒物对于呼吸系统细胞有毒性,RTCA法可以应用于雾霾细颗粒物体外毒性检测。Objective To detect the cytotoxicity of fine particulate matter treated on human bronchial epithelial cells（ BEAS-2B） and human embryonic lung fibroblasts（ MRC-5） by using the method of real time cell analyze（ RTCA） and Alamar Blue.Methods Seed the BEAS-2B cells and MRC-5 cells in the 16-well plates for RTCA analyzer and commonly 96-well plates respectively. After 24 hours,the BEAS-2B cells and the MRC-5 cells were treated by the fine particulate matter respectively in different doses（ 2500,1250,625,312. 5,156. 25 and 78. 125 μg / m L）. Calculating the IC50 values which determined by the two methods on different times（ 3,12,24,36 and 48 h）. Using paired t test by SPSS 16. 0 analyse the IC50 values from two methods and estimate the correlation. Results There was correlation between the two methods,but there was no obvious difference between the two results by RTCA and Alamar Blue. Conclusion There is certain cytotoxicity of fine particulate matter. At the same time,the RTCA real-time monitoring technology can be used on determination of cytotoxicity in vitro.

关 键 词：雾霾细颗粒物 呼吸系统 细胞毒性 实时细胞分析法 Alamar Blue法 

分 类 号：R122.1[医药卫生—环境卫生学]