基于Luminex液相芯片技术的NNK和AαC体外免疫毒性研究  

Evaluation of immunotoxicity of NNK and AαC in vitro by Luminex liquid chip technology

作　　者：赵俊伟[1] 杨陟华[2] 李翔[1] 谢复炜[1] 潘秀颉[2] 朱茂祥[2] 刘兴余[3] 尚平平[1] 康彧[1] 赵阁[1] 聂聪[1] 刘惠民[1] 谢剑平[1] 

机构地区：[1]中国烟草总公司郑州烟草研究院烟草化学重点实验室,河南郑州450001 [2]军事医学科学院放射与辐射医学研究所 [3]上海烟草集团北京卷烟厂

出　　处：《环境与健康杂志》2015年第11期958-962,共5页Journal of Environment and Health

基　　金：国家科技支撑计划项目(2012BAK01B03);郑州烟草研究院院长科技发展基金项目(322014CA0350);郑州烟草研究院科技项目(322013CZ0600)

摘　　要：目的了解4-(甲基亚硝胺基)-1-(3-吡啶)-1-丁酮(NNK)和2-氨基-9H-吡啶并[2,3-b]吲哚(AαC)的体外免疫毒性特征。方法分别以0、12.5、25、50、100、150和200μg/ml NNK染毒小鼠淋巴瘤细胞(EL-4细胞)24 h,以0、2.5、5、7.5、10、12.5和15μg/ml AαC染毒小鼠巨噬细胞(Ana-1细胞)24 h。采用CCK-8法检测细胞存活率,以Luminex液相芯片技术检测细胞上清液中与免疫相关的20种细胞因子分泌量,以BCA蛋白检测方法校正细胞因子的结果。结果 NNK染毒EL-4细胞后,白介素(IL)-10、IL-17、IL-1β和肿瘤坏死因子-α(TNF-α)含量随染毒剂量的增加而升高,粒细胞-巨噬细胞集落刺激因子(GM-CSF)含量随染毒剂量的增加而降低,且均存在剂量-效应关系;碱性成纤维细胞生长因子(FGF-basic)、γ-干扰素(IFN-γ)、IL-1α、IL-2、IL-4、IL-5、IL-6、IL-12、IL-13、干扰素诱导蛋白-10(IP-10)、角质细胞诱导因子(KC)、单核细胞趋化蛋白-1(MCP-1)、γ-干扰素诱导性单核因子趋化因子(MIG)、巨噬细胞炎症蛋白-1α(MIP-1α)和血管内皮生长因子(VEGF)含量无明显变化。AαC染毒Ana-1细胞后,IL-5、MCP-1、MIP-1α、TNF-α和VEGF含量随染毒剂量的增加而降低,其余15种细胞因子无明显变化。结论 NNK和AαC可以产生体外免疫毒性。Objective To determine immunotoxicity of NNK and AαC in vitro. Methods The exposure experiment of EL-4cells to NNK with the concentrations of 0, 12.5, 25, 50, 100, 150 and 200 μg/ml were conducted. The exposure experiment of Ana-1 cells to AαC with the concentrations of 0, 2.5, 5, 7.5, 10, 12.5 and 15 μg/ml were conducted. Cell viability was measured by CCK-8 assay. The levels of 20 cytokines in the culture media were detected by Luminex liquid chip.Concentration of cell protein in culture media was measured by trace bicinchoninic acid（BCA） method to correct cytokines.Results For EL-4 cells, NNK significantly induced IL-10, IL-17, IL-1β and TNF-α production in a concentration-dependent manner,whereas GM-CSF was inhibited in a concentration-dependent manner. The levels of FGF-basic, IFN-γ, IL-1α, IL-2,IL-4, IL-5, IL-6, IL-12, IL-13, IP-10, KC, MCP-1, MIG, MIP-1α and VEGF in EL-4 cells did not show significantly difference between exposure group and control（P〈0.05）. For Ana-1 cells,AαC significantly inhibited IL-5, MCP-1, MIP-1α,TNF-α and VEGF production in a concentration-dependent manner. FGF-basic, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-4,IL-6, IL-10, IL-12, IL-13, IL-17, IP-10, KC and MIG in Ana-1 cells did not show significantly difference between exposure group and control（P〉0.05）. Conclusion NNK and AαC are of immunotoxicity in vitro.

关 键 词：Luminex液相芯片 NNK AαC EL-4细胞 Ana-1细胞 细胞因子 

分 类 号：R994.6[医药卫生—毒理学]