Effects of Evodiamine and Rutecarpine on Human CYP 3A4 through PXR and CAR Pathway
OBJECTIVE:To study the effects of evodiamine and rutecarpine on the activity of cytochrome CYP 3A4 through pregnane X receptor (PXR) and constitutive androstane receptor (CAR) pathway.METHODS:PXR plasmid and the CYP3A4 report plasmid or CAR plasmid and the CYP3A4 report plasmid were transiently transfected into the LS174T cell,and the evodiamine and rutecarpine 2.5,10 and 40 μmol/L were incubated with the LS174T cell.The activities of luciferase were determined with CYP3A4 report gene.Primary hepatocytes were isolated from mice by two steps perfusion method and then treated with evodiamine and rutecarpine 2.5,10 and 40 μtmol/L.CYP3A11 mRNA expression and enzymatic activity of primary hepatocytes were measured using real-time PCR and LC-MS/MS.RESULTS:The evodiamine 10 and 40 μtmol/L could significantly enhanced 3.41 and 4.15 folds of luciferase activity in CYP3A4 report gene through PXR pathway.Rutecarpine 10 and 40 μmol/L could reduce the effect of rifampicin on PXR,decreasing by 37.7％ and 45.34％,respectively.Rutecarpine could reduce 29％ induction of CAR caused by CITCO (positive activator of CAR).After exposed to evodiamine 10,40 μmol/L,the expression of CYP3A11 mRNA was significantly 2.67 and 3.80 fold increased,respectively; the expressions of CYP3A11 mRNA in 35％ primary hepatocytes were significantly down-regulated by rutecarpine 40 μmol/L.Evodiamine 10,40 μmol/L increased 3.63 and 3.01 folds of CYP3A11 enzyme activity significantly; after exposed to rutecarpine 40 μmol/L,CYP3A4 enzyme activity was decreased by 34.54％ significantly.CONCLUSIONS:Evodiamine can significantly enhance the CYP3A4 luciferase,mRNA expression and enzyme activity through PXR mediated pathway.Rutecarpine can significantly inhibit the CYP3A4 luciferase,RNA expression and enzyme activity through PXR/CAR mediated pathway.
EvodiamineRutecarpineDrug interactionPregnane X receptorConstitutive androstane receptorCYP3A4
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