离体检测方法预测三氯乙烯致敏性  

Predicting the sensitizing potency of trichloroethylene by in vitro test

作　　者：曾丽海[1] 郑杰蔚 谢植伟[1] 钟怡洲 李国樑[1] 梁博萱 殷霄[1] 赖关朝[1] 黄振烈[1] ZENG Lihai;ZHENG Jiewei;XIE Zhiwei;ZHONG Yizhou;LI Guoliang;LIANG Boxuan;YIN Xiao;LAI Guanchao;HUANG Zhenlie(Guangdong Province Hospital for Occupational Diseases Prevention and Treatmen;Guangdong Provincial Key Laboratory of Occupational Disease Prevention and Treatment, Guangzhou, Guangdong 510300, Chin)

机构地区：[1]广东省职业病防治院广东省职业病防治重点实验室,广东广州510300

出　　处：《中国职业医学》2018年第3期279-284,共6页China Occupational Medicine

基　　金：国家科技支撑计划项目(2014BAI12B01);国家临床重点专科建设项目(2011-09);广东省医学科学技术研究基金(A2015139);广州市科技计划项目(201707010405);广东省职业病防治重点实验室(2017B030314152);广东省化学中毒与核辐射突发事件医学救援应急技术研究中心(2012A032100002)

摘　　要：目的采用人急性单核细胞白血病细胞系的THP-1细胞预测三氯乙烯(TCE)的致敏性及最佳的致敏剂量。方法离体培养THP-1细胞,以不同浓度2,4-二硝基氯苯(DNCB)、十二烷基硫酸钠(SDS)、叔丁基对苯二酚(tBHQ)和TCE与THP-1细胞共孵育24 h,采用流式细胞仪检测细胞表面标志物分化抗原(CD)86和CD54的表达,确定致敏检测的最佳剂量范围,并以相对荧光强度(RFI,CD86)≥150和RFI(CD54)≥200为标准,预测TCE的致敏性及最佳致敏剂量。结果采用THP-1细胞评价致敏性的合适浓度范围可能为细胞相对存活率在75.0%~100.0%时的浓度,DNCB在浓度为20.83、25.00和30.00μmol/L时,tBHQ在浓度为5.80μmol/L时,TCE在浓度为8.33、10.00和12.00 mmol/L时,均具有致敏性;SDS为非致敏物。TCE浓度为8.33 mmol/L时,CD86和CD54表达量最大,为最佳致敏剂量。结论 TCE的最佳致敏剂量为8.33 mmol/L,可为后续TCE致敏毒性通路的研究提供剂量设计的依据。Objective To predict the sensitizing potency and optimal sensitization dose of trichloroethylene（ TCE） by an in vitro skin sensitization test on a human acute monocytosis cell line（ THP-1）.Methods THP-1 cells were cultured in vitro and exposed to 2,4-dinitrochlorobenzene（ DNCB）,sodium dodecyl sulfate（ SDS）,tert-butylhydroquinone（ tBHQ）and TCE for 24 hours.Flow cytometry was used to detect the expression of cell surface marker such as cluster of differentiation（ CD） 86 and CD54,and the optimal dose range for sensitization detection was determined.With the relative fluorescence intensity（ RFI）,CD86 ≥ 150 and CD54 ≥ 200 as the standard,the sensitizing potency and optimal sensitization dose of TCE were predicted.Results The concentration range of reagents for sensitization test on THP-1 cells was the dose range at which the relative cell survival rate reached 75.0%-100.0%.DNCB at the doses of 20.83,25.00 and 30.00 μmol/L,tBHQ at the dose of 5.80 μmol/L,TCE at the doses of 8.33,10.00 and 12.00 mmol/L,can cause sensitivity.SDS was recognized as a negative sensitizer.The expression of CD86 and CD54 was the highest when the concentration of TCE was 8.33 mmol/L,which was considered as the best sensitization dose.Conclusion The optimum sensitization dose of TCE is 8.33 mmol/L,which can provide the basis for dose design in future study of TCE sensitization pathways.

关 键 词：三氯乙烯 皮肤致敏 THP-1细胞 离体实验 分化抗原 

分 类 号：R114[医药卫生—卫生毒理学] R135.7[医药卫生—公共卫生与预防医学]