摘要
目的探讨三氯乙烯(TCE)对体外诱导人未成熟树突状细胞(iDC)的细胞毒性作用。方法免疫磁珠分离人外周血单核细胞,应用重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)和白细胞介素-4(rhIL-4)诱导获得iDC,细胞形态学观察和流式细胞仪(FACS)检测细胞表面标记鉴定。给予TCE处理进行剂量-效应和时间-效应分析,采用Cell Counting Kit-8(CCK-8)检测细胞存活率。结果诱导获得的iDC呈悬浮状态,部分细胞聚集成簇,大部分细胞呈圆形,部分细胞的胞浆存在少量不明显的毛刺样树突伸出。细胞表面标志分子表达阳性率CD14为0.35%,CD1a为85.82%,HLA-DR为50.34%,CD40为23.20%,CD80为9.65%,CD83为25.73%,CD86为33.67%,获得的细胞具有iDC的特征性表型。TCE不同剂量(0.5、1.0、5.0、10.0 mmol/L)处理组随作用剂量增加,细胞数呈不同程度减少,细胞皱缩现象逐渐增多,原本成簇的iDC随TCE作用剂量的增加而逐渐变松散以至散开;TCE不同剂量(1.0、5.0、10.0 mmol/L)处理组作用不同时间(24、48 h),细胞存活率随染毒时间的延长而降低,差异有统计学意义(P<0.05)。结论 TCE具有人未成熟DC细胞的毒性效应。
Objective To study the cytotoxicity of trichloroethylene( TCE) on dendritic cells( DC). Methods Isolated human peripheral blood monocytes( PBMC) from healthy donors using magnetic activated cell sorting( MACS) technique were induced with rhGM-CSF and rhIL-4 to get immature dendritic cell( iDC),which were identified both using by detecting the surface markers with cell morphological observation and flow cytometry instrument( FACS) to detect cell surface marker. After treatment of TCE,the cells,Counting Kit-8( CCK-8) was used conduced to evaluate the cell viability of iDC treated by TCE and analyze its dose-effect and time-course studies effect relationships. Results It was showed that the induced iDC growed in a suspended status and some of them were clustered aggregated into clumps,majority with typical morphology round shape,the cytoplasm in some cells were found a little bit out of a small inconspicuous burr like dendrites. The cell surface marker expression positive rate were: There was high level expression simultaneously in CD1a( 85. 82%), HLA-DR( 50. 34%), and low level in CD14( 0. 35%), CD40( 23. 20%), CD80( 9. 65%), CD83( 25. 73%), CD86( 33. 67%),respectively. The number of iDCs showed decreasing trend in TCE treated group with TCE dose,the clustering tendency was gradually reduced,and the viability of iDC was significantly decreased too,with dose-and time-depended patterns( P <0. 05). Conclusion It was shown that there was some cytotoxic effect of TCE on human immature DC.
出处
《中国工业医学杂志》
CAS
北大核心
2014年第1期6-9,共4页
Chinese Journal of Industrial Medicine
基金
广东省医学科研基金(A2010057)
广东省科技计划项目(2010B050700026)
关键词
三氯乙烯
树突状细胞
细胞毒性
trichloroethylene(TCE)
dendritic cell(DC)
cytotoxicity
