基于ARE-Nrf2 的皮肤致敏体外检测方法及其在样品检测中的应用
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(1.上海海关动植物与食品检验检疫技术中心,上海 200135; 2.上海家化联合股份有限公司启初研究中心,上海 200082; 3.上海国际旅行卫生保健中心,上海 200335)

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R-33

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An in vitro skin sensitization testing method based on ARE-Nrf2 and its application in sample test
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(1. Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs Distric,Shanghai 200135, China. 2. Giving Resserch Center, Shanghai Jahwa United Co. Ltd.,Shanghai 200082. 3. Shanghai International Travel Healthcare Center, Shanghai 200335)

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    摘要:

    目的 建立基于ARE-Nrf2 荧光素酶的皮肤致敏体外检测方法,并对化学品及混合物样品进行皮肤致敏性检测。方法 构建含有选择性质粒的稳转HaCaT 细胞株(DSens),不同浓度受试物与细胞共孵育48 h,通过荧光的读取检测细胞荧光素酶的表达量,从而判断物质的致敏潜力。对16 种已知皮肤致敏化学物质和6 种混合物样品进行皮肤致敏性预测。结果 使用DSens 的方法可准确区分16 种参考物质的皮肤致敏性,6 种混合物样品中,5 种预测结果与其他检测方法预测结果一致,另一种未知样品预测为阳性。结论 DSens 体外检测方法可以代替部分动物测试,用于可溶性皮肤致敏物质的预测。

    Abstract:

    Objective To establish a method for an in vitro test for skin sensitization based on ARE-Nrf2 (antioxidant responsive element - NF-E2-related factor 2) luciferase, and using this test to assess the sensitizing potency of chemicals and sample mixtures. Methods A stable transfection of the HaCaT cell line containing a selective plasmid encoding AKR1C2-ARE fused to the fluorescent luciferase was constructed (DSens), and the cells were incubated with samples for 48 hours at different concentrations. The expression of luciferase was measured by fluorescence detection. Skin sensitization was predicted for 16 known skin sensitizers and six mixtures. Results Sixteen reference substances were accurately distinguished by DSens. Among the six sample mixtures, the prediction result of five mixtures were consistent with the result of other detection method , and another unknown sample was predicted to be positive. Conclusions The DSens in vitro testing method may be a useful replacement for some animal tests used to predict sensitivity to soluble skin products.

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刘春凤,张丽婷,李小林,施镇,张子龙,李健,陈田,邱璐.基于ARE-Nrf2 的皮肤致敏体外检测方法及其在样品检测中的应用[J].中国比较医学杂志,2019,29(12):23~30.

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  • 收稿日期:2019-06-12
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  • 在线发布日期: 2020-01-13
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