Establishment of a PPAR-α Agonist Screening System Base on Report Gene

OBJECTIVE To establish a PPAR-α agonist screening system using a PPAR chimera system and double luciferase reporter gene assay method. METHODS pcDNA-hPPAR-α, pGL3-PPRE-luc and pRL-CMV (0.67 μg each) were Co-transfected into 293T cell seeded in 96 wells plate, fenofibrate was added into different groups after 6 hours transfection, respectively, up to terminal concentration of 0, 0.1, 1, 10, 50 μmol·L^-1 each. Two luciferase active were determined using Dual-Glo lucifarase assay system after 24 hours incubation, and relative luciferase intensive was used to express induction activity. RESULTS There is significant deviation between the 0.1, 1, 10, 50 μmol·L^-1 concentration point of fenobibrate, the relative induction activity of them were 0.82, 1.29, 1.72, 1.94. CONCLUSION A PPAR-α agonist screening system base on PPAR chimera system and double luciferase reporter gene assay method is established successfully.