中药组分丹参素对OATP1B1摄取瑞舒伐他汀的影响

Impact of Danshensu,a traditional Chinese medicine,on the transport of rosuvastatin mediated by OATP1B1

目的:在HepG2细胞中研究中药组分丹参素对SLCO1B1 mRNA及OATP1B1表达的影响,及体外对OATP1B1摄取瑞舒伐他汀的影响。方法:用RT-PCR法检测SLCO1B1 mRNA的表达,Western blot法检测OATP1B1的表达;建立LCMS分析方法检测HepG2细胞中瑞舒伐他汀的含量。以利福平(OATP1B1诱导剂)为探针开展平行对照实验,将系列浓度丹参素(20、40、80、120μmol·L^(-1))、利福平(5、10、25、50、100μmol·L^(-1))分别作用于HepG2细胞48 h,研究丹参素对SLCO1B1mRNA及OATP1B1表达的影响。通过摄取实验分别研究丹参素浓度(10、20、40、80、120μmol·L^(-1))及诱导时间(0、12、24、36、48 h)对HepG2细胞中瑞舒伐他汀摄取转运的影响。结果:不同质量浓度丹参素处理HepG2细胞48 h后,明显上调SLCO1B1 mRNA和OATP1B1表达水平,并随浓度增加而增强。在不同时间处理丹参素诱导下,12、24 h时瑞舒伐他汀摄取量分别减少22.02%和12.20%,而36、48 h时瑞舒伐他汀摄取量分别增加1.83%和85.39%。进一步考察发现,在48 h后用10、20μmol·L^(-1)丹参素处理可使瑞舒伐他汀摄取量分别减少6.89%、22.10%,而40、80、120μmol·L^(-1)丹参素则分别使其增加了1.23%、79.93%、87.64%。结论:丹参素能够诱导HepG2细胞中SLCO1B1 mRNA及OATP1B1的表达,并影响OATP1B1转运功能。丹参素低浓度短时间处理抑制HepG2细胞中OATP1B1对瑞舒伐他汀的摄取转运能力,高浓度长时间处理则诱导HepG2细胞中OATP1B1对瑞舒伐他汀的摄取转运能力。

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、120μmol·L^(-1))and rifampicin(5、10、25、50、100μmol·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、120μmol·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 120μmol·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 120μmol·L^(-1)for 48 h,uptake experiments were conducted with Danshensu as a substrate at a concentration of 58.55μmol·L^(-1)and rosuvastatin at a concentration of 30μmol·L^(-1).As compared with control group,Danshensu at concentrations of 10 and 20μmol·L^(-1)lowered rosuvastatin uptake by 6.89%and 22.10%while Danshensu at concentrations of 40/80/120μM 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.