Establishment of hPXR mediated reporter gene model for UGT1A1 and its application / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop hPXR mediated reporter gene model for UGT1A1, and use it as an in vitro model for determination of induction activity toward UGT1A1 of various commonly used traditional Chinese medicines (TCMs) extracts. METHODS: The distal and proximal promoters of UGT1A1 were amplified from human genetic DNA, and were cloned into pGL3 vector as pGL3-PXRE plasmid, which was then cotransfected into HepG2 cells with hPXR expression plasmid to establish the reporter gene model. Then the model was applied to determine the induction activity toward UGT1 A1 of various commonly used TCMs extracts. RESULTS: The promoters of UGT1A1 were successfully cloned into pGL3 vector to form pGL3-PXRE recombinant plasmid, and the reporter gene model was successfully established. Nine kinds of TCMs extracts were investigated, and three were found to activate hPXR and therefore have the potential to induce UGT1A1. CONCLUSION: The model established is an effective method for determination of activators of hPXR and potential inducers of UGT1A1 in vitro. Copyright 2012 by the Chinese Pharmaceutical Association.