熊果酸对基于OATP1B1与BCRP介导瑞舒伐他汀肝胆转运的影响及其机制研究

Chinese herbal medicines such as hawthorn, salvia etc are usually combined with statins so as to treat cardiovascular diseases more effective . Chinese herbal medicines contain many kinds of active components, which may have drug-drug interactions with statins. In preliminary studies，the result showed that ursolic acid significantly affected pharmacokinetics of rosuvastatin in rats, but mechanism of the effect was unclear at present.This subject, based on preliminary studies, taking OAPT1B1 and BCRP-mediated transport of rosuvastatin in hepatobiliary as the breakthrough point, will explore the effect of ursolic acid on OAPT1B1 and BCRP-mediated transport of rosuvastatin by establishing animal model, rat primary hepatocytes model and transgenic cell model. Given the genetic polymorphisms of OATP1B1 and BCRP, simultaneously, this subject will further explore the effect of ursolic acid on OATP1B1(521T>C) and BCRP(421C>A)-mediated transport of rosuvast-.atin.This subject aims to explore the effect and mechanism that ursolic acid affect OATP1B1 and BCRP-mediated transport of rosuvastatin in hepatobiliary at animal, cell and molecular level, elucidate the mechanism that ursolic acid affected the pharmacokinetics of rosuvastatin and provide new research method and idea to explore the mechanism of drug interaction between Chinese traditional medicine active components and chemical drugs.

中草药如山楂、丹参等常与他汀药物联合用药以更有效的治疗心血管疾病。中草药含有多种活性成分，存在与他汀药物发生药物相互作用的可能。前期实验研究结果显示熊果酸显著影响瑞舒伐他汀在大鼠体内的药动学，但该影响机制目前尚不明确。本课题拟在前期研究基础上，以OAPT1B1与BCRP介导瑞舒伐他汀肝胆转运为切入点，通过建立整体动物模型、大鼠原代肝细胞模型与转基因细胞模型，研究熊果酸对OATP1B1与BCRP介导瑞舒伐他汀转运的影响及机制；鉴于OATP1B1与BCRP的基因多态性，将同时进一步探讨熊果酸对突变型OATP1B1（521T>C）与BCRP（421C>A）介导瑞舒伐他汀转运的影响。本课题旨在动物、细胞与分子水平上探索熊果酸对OATP1B1与BCRP介导瑞舒伐他汀肝胆转运的影响及其机制；阐明熊果酸对瑞舒伐他汀药动学行为的影响机制，为探索中药活性成分与化学药物间相互作用的机制研究提供新的方法与思路。

中药成分可能作用于不同药物转运体，对药物转运体介导其它药物的转运功能产生作用，最终影响药动学过程。前期实验结果显示熊果酸显著影响瑞舒伐他汀在大鼠体内的药动学，但该影响机制目前尚不明确。本课题拟以OAPT1B1与BCRP介导瑞舒伐他汀肝胆转运为切入点，通过建立整体动物模型、大鼠原代肝细胞模型与转基因细胞模型，研究熊果酸对OATP1B1与BCRP介导瑞舒伐他汀转运的影响及机制；鉴于OATP1B1与BCRP的基因多态性，将同时进一步探讨熊果酸对突变型OATP1B1（521T>C）与BCRP（421C>A）介导瑞舒伐他汀转运的影响。研究结果显示合用熊果酸与Ko143均可显著影响大鼠体内瑞舒伐他汀的药动学参数Cmax(mg.L-1)（2.23±0.22 vs 3.86±0.29vs4.18±0.18），AUC0-t (mg.L*min-1)( 31.99±1.93vs53.14±2.08vs62.72±6.34)；当熊果酸浓度为4，8，16uM时，肝细胞摄取瑞舒伐他汀的量分别减少了1.70 ± 0.94, 47.58 ± 1.80 和71.16 ± 0.19 %，抑制参数Ki为10.88 ± 0.29 uM；利用转基因技术建立了稳定表达OATP1B1与BCRP的细胞模型(OATP1B1*1a-HEK 293T , OATP1B1*5-HEK 293T，BCRP421CC –MDCK, BCRP421AA –MDCK)，并应用于本课题的细胞动力学研究，结果显示当熊果酸的浓度为1.8和18uM时，OATP1B1*1a 与OATP1B1*5介导瑞舒伐他汀的转运分别减少了34.60 ± 2.99 %和 66.08 ± 1.83 %与34.27 ± 7.08 % 和 66.95 ± 1.14 %，IC50 分别为 6.25 ± 0.42 和6.07 ± 0.57 uM；当熊果酸浓度为25uM时，与对照组相比，瑞舒伐他汀在BCRP421CC–MDCK与BCRP421AA –MDCK细胞单层模型中的Efflux ratio发生变化（4.16±0.3vs1.78±0.16; 4.12±0.36vs2.12±0.36）。本课题旨在动物、细胞与分子水平上阐明熊果酸对瑞舒伐他汀药动学行为的影响机制，为探索中药活性成分与化学药物间相互作用的机制研究提供新的方法与思路。研究结果发表3篇外文文章。