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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R-33

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    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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History
  • Received:June 12,2019
  • Revised:
  • Adopted:
  • Online: January 13,2020
  • Published: