三氯乙烯的皮肤致敏性及对小鼠淋巴结CD4^+T细胞活化的影响  

The skin sensitization and effect on the activation of lymph nodes CD4^+T cells of trichloroethylene

作　　者：杨进涛[1] 赵燕[1] 唐秋琼[1] 蒋建军[1] 魏雪涛[1] 尚兰琴[1] 郝卫东[1] 

机构地区：[1]北京大学公共卫生学院食品安全毒理学研究与评价北京市重点实验室,北京100191

出　　处：《毒理学杂志》2012年第4期270-273,共4页Journal of Toxicology

基　　金：国家重大新药创制专项(2009ZX09301-010)

摘　　要：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 Using LLNA BrdU-LLNA method to assess the skin sensitization and effect on the activation of CD4＋ T cells of trichloroethylene. Methods C57 mice were randomly divided into AOO vehicle control group, TCE tested groups and hexyl cinnamie aldehyde positive control group. 25 μL of the appropriate dilution of the test substance, the vehicle alone, or the PC were applied to the dorsum of each ear on day I , 2 and 3. 0. 5ml of BrdU （10mg/ml） solution were injected inter-peritoneally to each mouse on clay 6. Approximately 24 hours after BrdU injection, the animals were humanely killed, the draining auricular lymph nodes were excised and single- cell suspensions were generated by gentle mechanical disaggregation through 200 micron-mesh gauze. The counts of lymph node cells and BrdU labeling index were measured and the SI was calculated for each tested chemicals. The percentage of CD44＋ CD62L- cells,IFN -γ＋ cells and Foxp3 ＋ cells in CD4＋ T cells in lymph nodes were detected with flow cytometer. Results The organ coefficient of lymph nodes increased in 80% TCE and 25% HCA groups. The BrdU labeling index of TCE tested groups were significant higher than that of vehicle control group, and had an apparent dose-response relationship. 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 respectively. The percentage of CD44＋ CD62L- cells,IFN-γ＋ ceils and Foxp3＋ cells in CD4＋ T cells of 80% TCE group were significant higher than that of vehicle control group. Conclusions BrdU- LLNA might be an useful method for evaluating the skin sensitization of chemicals. Trichloroethylene might be a skin sensitizer. Trichloroethylene could activate CD4＋T cells of lymph nodes, and increase the percent of Thl cells and Treg cells in CD4 ＋T cells.

关 键 词：局部淋巴结试验 三氯乙烯 皮肤致敏性 

分 类 号：R99[医药卫生—毒理学]