Glue C, Millner A, Bodtger U, Jinquan T, Poulsen L K
Laboratory of Medical Allergology, Allergy Unit, National University Hospital, Blegdamsvej 9, Dept. 7542, DK-2100 Copenhagen, Denmark.
Toxicol In Vitro. 2002 Dec;16(6):657-62. doi: 10.1016/s0887-2333(02)00082-6.
It has recently been shown that plasticizers are present in indoor air dust, which may lead to human exposure via the inhalation route. Moreover, studies have indicated that plasticizers may possess adjuvant effects increasing the health damaging potential of allergens. The aim of this study was to investigate the in vitro effect of metabolites of phthalate plastisizers, such as whether an adjuvant effect is paralleled by changes of the cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells (PBMCs) from allergics and non-allergics. The toxicity monitored by cell viability was determined by incubating THP-1 cells with a 10-fold dilution series of monophthalates for 24 h. At different points in time cytokine expression (IL-1beta, IL-6, IL-12alpha (p35)) in THP-1 cells incubated with non-toxic concentrations of monophthalate (2-20 microg/ml)+/-LPS (1 microg/ml) were determined using Quantitative Competitive RT-PCR. PBMCs from allergics and non-allergics were incubated with monophthalate 220 microg/ml) for up to 48 h and cytokine expression (IL-4, IL-5, IFN-gamma) was measured using real-time PCR. The cytotoxic level of monophthalates is 20-200 microg/ml, depending on the individual monophthalate. There seems to be a correlation between increasing side-chain length and toxicity. Monophthalates did not induce changes in cytokine expression in THP-1 cells, though there is an increase when co-incubating with LPS. Cytokine expression in PBMC seems virtually unchanged when co-incubated with monophthalate, though mono-n-butyl phthalate (MBUP) tends to increase the level of IL-4 in PBMCs from allergic individuals. The two cellular models demonstrated the dynamics of regulated cytokine mRNA and are applicable for in vitro immunotoxicological investigations. The results regarding monophthalates suggest these to have a limited effect on cytokine expression in the monocytic cell line THP-1 and weak effect on cytokine expression in PBMCs from allergic and non-allergic individuals.
最近研究表明，室内空气灰尘中存在增塑剂，这可能导致人类通过吸入途径接触到增塑剂。此外，研究表明增塑剂可能具有佐剂效应，会增加过敏原对健康的损害潜力。本研究的目的是调查邻苯二甲酸酯类增塑剂代谢产物的体外效应，例如在单核细胞系THP-1以及过敏者和非过敏者的外周血单个核细胞（PBMC）中，佐剂效应是否与细胞因子表达的变化同时出现。通过用10倍稀释系列的单邻苯二甲酸酯孵育THP-1细胞24小时，测定细胞活力来监测毒性。在不同时间点，使用定量竞争RT-PCR测定用无毒浓度的单邻苯二甲酸酯（2 - 20微克/毫升）±脂多糖（1微克/毫升）孵育的THP-1细胞中的细胞因子表达（IL-1β、IL-6、IL-12α（p35））。将过敏者和非过敏者的PBMC与单邻苯二甲酸酯（2 - 20微克/毫升）孵育长达48小时，使用实时PCR测量细胞因子表达（IL-4、IL-5、IFN-γ）。单邻苯二甲酸酯的细胞毒性水平为20 - 200微克/毫升，具体取决于单个单邻苯二甲酸酯。侧链长度增加与毒性之间似乎存在相关性。单邻苯二甲酸酯不会诱导THP-1细胞中细胞因子表达的变化，不过与脂多糖共同孵育时会增加。与单邻苯二甲酸酯共同孵育时，PBMC中的细胞因子表达实际上没有变化，不过邻苯二甲酸单丁酯（MBUP）倾向于增加过敏个体PBMC中IL-4的水平。这两种细胞模型展示了细胞因子mRNA表达调控的动态过程，适用于体外免疫毒理学研究。关于单邻苯二甲酸酯的结果表明，它们对单核细胞系THP-1中的细胞因子表达影响有限，对过敏和非过敏个体PBMC中的细胞因子表达影响较弱。
