基于Luminex液相芯片技术的NNK和AαC体外免疫毒性研究

发布时间：2018-03-16 12:46

  本文选题：Luminex液相芯片　切入点：NNK　出处：《环境与健康杂志》2015年11期 　论文类型：期刊论文

【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。
[Abstract]:Objective to investigate the characteristics of in vitro immunotoxicity of 4-( methylnitrosamine-3-pyridine-3-pyridine-1-butanone) and 2-amino-9H-pyridyl [2Amino-9H-pyridine] indole A 伪 C. Methods mice lymphoma cell line EL-4 cells were exposed to 0.12.5rml NNK for 24 h and 2.57.51010g / ml to 12.5 and 15 渭 g / ml A 伪 -C, respectively. The survival rate of murine macrophages, Ana-1 cells, was determined by CCK-8 assay for 24 h. The levels of immune-related cytokines in supernatant of cells were detected by Luminex liquid chip technique, and the results of cytokines were corrected by BCA protein assay. Results after NNK was exposed to EL-4 cells, The contents of IL-17 尾 and TNF- 伪 increased with the increase of exposure dose, and the content of granulocyte-macrophage colony stimulating factor (GM-CSF) decreased with the increase of dose, and there was a dose-effect relationship between the contents of IL-17 尾 and TNF- 伪. Basic fibroblast growth factor FGF-basicus, IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IFN- 纬 -IL-2-, IL-5, IL-6, IL-6, IL-12, Interferon-inducible protein (-10), keratinocyte inducible factor (KCU), monocyte chemoattractant protein (MCP-1), interferon- 纬 -inducible monokine chemokine (IFN- 纬), macrophage inflammatory protein 1 伪 (MIP-1 伪) and intravascular inflammation protein (MIP-1 伪). There was no significant change in the content of skin growth factor (VEGF). The contents of TNF- 伪 and VEGF decreased with the increase of dose of Ana-1 cells. Conclusion NNK and A 伪 C can produce in vitro immunotoxicity.
【作者单位】： 中国烟草总公司郑州烟草研究院烟草化学重点实验室;军事医学科学院放射与辐射医学研究所;上海烟草集团北京卷烟厂;
【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)
【分类号】：R994
，

本文编号：1619976