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目的通过局部淋巴结试验BrdU-ELISA法研究三氯乙烯(TCE)的皮肤致敏性及对小鼠淋巴结CD4+T细胞活化的影响。方法 28只雌性C57小鼠随机分7组,分别为AOO溶剂对照组、5%TCE组、10%TCE组、20%TCE组、40%TCE组、80%TCE组和25%己基肉桂醛(HCA)阳性对照组,实验第1、2、3天分别于小鼠双耳耳背部涂抹25μl受试物,第6天经腹腔注射0.5ml 10mg/ml BrdU溶液,第7天处死小鼠,取双侧耳后淋巴结,进行淋巴细胞计数和BrdU含量测定,比较各组小鼠BrdU标记指数和刺激指数(SI)的差异。同时用流式细胞仪检测淋巴结CD4+T细胞中CD44+CD62L-细胞、IFN-γ+细胞(Th1细胞)和Foxp3+细胞(Treg细胞)比例。结果 80%TCE组和25%HCA组与AOO溶剂对照组相比,小鼠淋巴结脏器系数和淋巴结淋巴细胞数显著增加;TCE各浓度组BrdU标记指数显著增高,具有剂量-反应关系,25%HCA组BrdU标记指数显著高于AOO溶剂对照组;5%TCE、10%TCE、20%TCE、40%TCE、80%TCE和25%HCA的刺激指数(SI)依次为:2.24、1.79、1.97、2.36、2.81和2.62。80%TCE组小鼠淋巴结CD4+T细胞中CD44+CD62L-细胞、IFN-γ+细胞和Foxp3+细胞比例依次为22.30%、3.90%和17.05%,均高于溶剂对照组。结论局部淋巴结试验BrdU-ELISA法是一种较好的评价化学物皮肤致敏性的替代试验方法,TCE是一种皮肤致敏物。TCE可以促进小鼠淋巴结CD4+T细胞活化,并提高CD4+T细胞中Th1细胞和Treg细胞比例。
Objective To study the skin sensitization of trichlorethylene (TCE) and its effect on the activation of CD4 + T lymphocytes in lymph nodes of mice by BrdU-ELISA assay. Methods Twenty-eight female C57 mice were randomly divided into 7 groups: AOO solvent control group, 5% TCE group, 10% TCE group, 20% TCE group, 40% TCE group, 80% TCE group and 25% hexylcinnamaldehyde HCA) positive control group. On day 1, 2, and 3 of the experiment, 25 μl of the test substance was smeared on the backs of the ears of mice. 0.5 ml of 10 mg / ml BrdU solution was injected intraperitoneally on the 6th day. Bilateral posterior lymph nodes, lymphocyte count and BrdU content were measured, and the differences of BrdU labeling index and stimulation index (SI) between the two groups were compared. Meanwhile, the proportion of CD44 + CD62L-, IFN- γ + cells (Th1 cells) and Foxp3 + cells (Treg cells) in lymph node CD4 + T cells were detected by flow cytometry. Results Compared with AOO solvent control group, the numbers of lymph node organ and lymph node lymphocytes in 80% TCE group and 25% HCA group increased significantly. The BrdU labeling index in each concentration group of TCE was significantly increased, with a dose-response relationship of 25% The BrdU labeling index of HCA group was significantly higher than that of AOO solvent control group. The stimulation index (SI) of 5% TCE, 10% TCE, 20% TCE, 40% TCE, 80% TCE and 25% HCA were 2.24,1.79,1.97 , 2.36, 2.81 and 2.62.80% respectively. The proportion of CD44 + CD62L-, IFN- γ + cells and Foxp3 + cells in lymphocytes of TCE group were 22.30%, 3.90% and 17.05% group. Conclusion The local lymph node test BrdU-ELISA method is a good alternative to evaluate the chemical skin sensitization alternative test method, TCE is a skin sensitizer. TCE can promote lymphocyte CD4 + T cell activation in mice and increase the proportion of Th1 cells and Treg cells in CD4 + T cells.