LLNA:BrdU-ELISA改良法的建立及其在化妆品安全性评价中的应用
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中青年基金项目(2013NC3)。


Establishment of modified LLNA:BrdU-ELISA and its application in the safety evaluation of cosmetics
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    摘要:

    目的 建立可快速检测化学物致敏性和刺激性的局部淋巴结试验(LLNA:BrdU-ELISA)改良法,并对化妆品产品进行评价。方法 3种化学物(2,4二硝基氯苯(DNCB)、丁子香酚、己基肉桂醛)和3种化妆品产品作为受试物,雌性BALB/c小鼠连续染毒3天,测量小鼠耳缘厚度,第5天腹腔注射BrdU,第6天称耳廓重,分离颌下淋巴结称重并制取单细胞悬液,用ELISA试剂盒检测淋巴细胞增殖。结果 DNCB(1.0%)、己基肉桂醛(25%、50%)和3号粉底霜引起耳肿胀和耳廓重显著增加(P < 0.05或P < 0.01),可能为刺激物,其他均无刺激性;3种化学物和3号粉底霜致敏检测阳性,其他均为阴性。结论 结合小鼠耳肿胀和耳廓重的LLNA:BrdU-ELISA改良法可较好地评价化学物和化妆品产品的致敏性和刺激性,有望在化妆品安全性评价中发挥重要作用。

    Abstract:

    Objective To establish a rapid modified Local lymph node assay (LLNA) for the sensitization and irritancy potential of chemicals, and evaluate cosmetics products. Methods 3 kinds of chemicals (2, 4-Dinitrochlorobenzene (DNCB), Eugenol and Hexyl cinnamic aldehyde) and 3 kinds of cosmetics were selected. The dorsal sides of both ears of female BALB/c mice were treated with 25μL/ear of the test solutions on 3 consecutive days, and the thickness of each ear was measured. BrdU solution was injected inter-peritoneally on day 5. On day 6, each auricle was weighed, the bilateral draining auricular lymph nodes were excised and weighed, then made into single cell suspension. BrdU was measured by ELISA using a commercial kit. Results Ear swelling and auricle weight of DNCB (1.0%), Eugenol (25% and 50%) and number 3 cream foundation increased significantly (P < 0.05 or P < 0.01), which may be considered as irritants. The others were negative. For LLNA, three chemicals and number 3 cream foundation pretended positive. Conclusion The combined LLNA/ear swelling/ear weight assay appears to be a reliable predictor of sensitization and irritancy potential, and is expected to play an important role in safety evaluation of cosmetics.

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胡培丽,罗飞亚,陈志蓉,邢书霞. LLNA:BrdU-ELISA改良法的建立及其在化妆品安全性评价中的应用[J].中国比较医学杂志,2014,24(9):23~27.

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  • 最后修改日期:2014-08-25
  • 在线发布日期: 2014-10-10
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