Abstract: Objective: To establish the Nrf2 signaling pathway activation reporting system with secreted Gaussia luciferase (Gluc) as report gene. Methods: The transcribed sequences based on CV-ARE and ML-ARE, Gluc as a reporter gene, were constructed. Alkaline phosphatase (AP) promoted by ?ML and CV promoter as an internal report, different numbers (1, 2, 4 or 8) of ARE were inserted into EcoRI---XhoI respectively. The activity of Gluc with different numbers of ARE were compared. Results: The fluorescence activity of Gluc with ML-8XARE and CV-8XARE was the highest.. The interference of ML-ARE to antioxidant was lower than CV-ARE. The ML-8XARE plasmid was transfect to HaCaT cells, then were treated by sulforaphane (SF) and tert butyl hydroquinone (TBHQ) or UV. The activity of Gluc was the highest at 6 hours after treated by SF and TBHQ and at 24 hours radiated by UV. Conclusion: The Nrf2 signaling pathway activation reporting system with secreted Gaussia luciferase (Gluc) as report gene was successfully constructed, which lay the foundation for the model of UV disease.

Key words: reporting system, Nrf2, Gluc