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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。
Aim To investigate the effect of 4- (methylnitrosamino) -1- (3-pyridyl) -1-butanone (NNK) and 2-amino-9H-pyrido [2,3- b] In Vitro Immunotoxicity. Methods Mouse lymphoma cells (EL-4 cells) were exposed to 0,2.5,25,50,100,150 and 200μg / ml NNK for 24 h, and treated with 0,2.5,5,7.5,10,12.5 and 15μg / ml AαC mouse macrophages (Ana-1 cells) 24 h. Cell viability was detected by CCK-8 assay, and 20 immune-related cytokines secreted by the supernatant of the cells were detected by Luminex liquid-phase microarray. The cytokine results were corrected by BCA protein assay. Results The levels of interleukin (IL) -10, IL-17, IL-1β and tumor necrosis factor-α (TNF-α) increased with the increase of the dose of NNK in EL-4 cells. The content of GM-CSF decreased with the increase of dose, and all had dose-effect relationship. The levels of FGF-basic, IFN-γ, , IL-1α, IL-2, IL-4, IL-5, IL-6, IL- 12, IL- 13, interferon- inducible protein-10 (IP- 10), keratinocyte-inducing factor The levels of MCP-1, MIG, MIP-1α and VEGF were measured by MTT assay. No significant changes. The levels of IL-5, MCP-1, MIP-1α, TNF-αand VEGF in AaC-treated Ana-1 cells decreased with the increase of the dose of AαC, while the other 15 kinds of cytokines did not change significantly. Conclusion NNK and AαC can produce immunotoxicity in vitro.