Impact of Danshensu,a traditional Chinese medicine,on the transport of rosuvastatin mediated by OATP1B1
OBJECTIVE To explore the impact of Danshensu on the expression of SLCO1B1 mRNA and OATP1B1 and elucidate the mechanism of Danshensu on the uptake of rosuvastatin by OATP1B1 in HepG2 cells.METHODS The expression level of SLCO1B1 mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR)and protein expression of OATP1B1 by Western blot.Liquid chromatography-mass spectrometry(LC-MS)analysis method was employed for detecting the content of rosuvastatin.Parallel control experiments were conducted with rifampicin as an inducer of OATP1B1.Different concentrations of Danshensu(20、40、80、120 μmol·L-1)and rifampicin(5、10、25、50、100 μmol·L-1)were separately applied for 48 h.The effect of Danshensu on SLCO1B1 mRNA and OATP1B1 expression was examined.The effects of different concentra-tions of Danshensu(10、20、40、80、120 μmol L-1)and induction durations(0、12、24、36、48 h)on uptake transport of rosuvastatin were investigated through intake experiments.RESULTS The application of Danshensu at 20 to 120 μmol·L-1 for 48 h signifi-cantly up-regulated the expressions of SLCO1B1 mRNA and OATP1B1 protein with a stronger induction effect observed at higher concentrations.Different induction durations(0-48 h)of Danshensu were applied for determining their impacts on the expression of OATP1B1.At 12/24 h,there was a significant reduction in uptake amount of rosuvastatin by 22.02％and 12.20％.The uptake amount of rosuvastatin spiked by 1.83％and 85.39％at 36/48 h.After an induced expression of OATP1B1 by Danshensu at concentrations of 0 to 120 μmol·L-1 for 48 h,uptake experiments were conducted with Danshensu as a substrate at a concentration of 58.55 μmol·L-1 and rosuvastatin at a concentration of 30 μmol·L-1.As compared with control group,Danshensu at concentrations of 10 and 20 μmol·L-1 lowered rosuvastatin uptake by 6.89％and 22.10％while Danshensu at concentrations of 40/80/120 μM boosted rosuvastatin uptake by 1.23％,79.93％and 87.64％respectively.CONCLUSION Danshensu may up-regulate the expressions of SLCO1B1 mRNA and OATP1B1 protein,thereby enhancing the uptake capacity of OATP1B1 in HepG2 cells.Short-term exposure to low concentrations suppresses the transport of rosuvastatin by OATP1B1,whereas long-term exposure to high concentrations induces the transport of rosuvastatin by OATP1B1.
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