淫羊藿苷元对临床重要药物转运体抑制作用研究

Inhibition effects of anhydroicaritin on important clinical drug transporters

目的研究淫羊藿苷元对临床重要转运体OAT1、OAT3、OATP1B1、OATP1B3、OCT1、OCT2、BCRP、BSEP和P-gp转运蛋白的抑制作用。方法应用过表达各转运体的细胞株,检测淫羊藿苷元(0、0.03、0.10、0.30、1.00、3.00、10.00μmol/L)对h OAT1介导的放射性同位素标记底物^(14)C-对氨基马尿酸(^(14)C-PAH)、h OAT3介导的3H-硫酸雌酮铵(3H-ES)、h OATP1B1介导的3H-ES、h OATP1B3介导的3H-雌二醇葡糖苷酸(3H-EG)、hOCT1介导的^(14)C-溴化四乙胺(^(14)C-TEA)、hOCT2介导的^(14)C-TEA、h BCRP介导的3H-ES、MDR1介导的3H-地高辛(3H-Digoxin)、以及h BSEP介导的3H-牛黄胆酸盐(3H-TCA)转运活性的影响,计算淫羊藿苷元对不同转运蛋白的抑制作用的半数抑制浓度(IC50)。结果与对照组比较,淫羊藿苷元0.3~10μmol/L浓度对OAT3转运活性发挥显著抑制作用(P<0.05),0.1~10μmol/L对P-gp有显著抑制作用;1~10μmol/L浓度对OATP1B3和BCRP转运活性有显著抑制作用(P<0.05);3~10μmol/L浓度对和OATP1B1转运活性有显著抑制作用(P<0.05);对OAT3和BCRP转运活性的IC50分别为4.97和8.15μmol/L;对OAT1B1、OATP1B3、OCT2和P-gp的IC50均大于10μmol/L,对OAT1、OCT1和BSEP转运活性无明显影响。结论淫羊藿苷元对药物转运体OAT3和BCRP的抑制作用相对较强,对OAT1B1、OATP1B3、OCT2和P-gp也具有一定的抑制作用,对OAT1、OCT1和BSEP无显著抑制作用。

Objective To research the inhibition of anhydroicaritin to the important clinical transporters including OAT1, OAT3, OATP1 B1, OATP1 B3, OCT1, OCT2, BCRP, BSEP, and P-gp. Method The transgenic cell lines overexpressing OAT1, OAT3, OATP1 B1, OATP1 B3, OCT1, OCT2, BCRP, BSEP, and P-gp were constructed, and the inhibition of anhydroicaritin（0, 0.03, 0.10, 0.30, 1.00, 3.00, and 10.00 μ mol/L） on the different transporters were evaluated by the effects of anhydroicaritin on transport activity of the radiolabeled substrates of ^14C-PAH, ^3H-ES, ^3H-ES, ^3H-EG, ^14C-TEA, ^14C-TEA, ^3H-ES, ^3H-Digoxin, 3 H-TCA mediated by h OAT1, h OAT3, OATP1 B, h OATP1 B3, h OCT1, h OCT2, h BCRP, MDR1,and h BSEP, respectively. The half maximal inhibitory concentration（IC50） of anhydroicartin to every transporters were calculated by the software Prism 5.0. Result Compared with control group, anhydroicaritin of 0.3 – 10 mol/L concentration had significant inhibitory effects on the transport activity of OAT3, BCRP, and OATP1 B3（P〈0.05）, and concentration of 0.1 - 10 and 1 - 10 μmol/L significantly inhibited the P-gp and OATP1 B1 transport activity（P〈0.05）. The IC50 of anhydroicaritin to OAT3 and BCRP transport activity was 4.97 and 8.15 mol/L, respectively, and IC50 of OAT1 B1, OATP1 B3, OCT2, and P-gp were more than 10 μmol/L, and with no obvious effects on the transport activity of OAT1, OCT1, and BSEP. Conclusion Anhydroicaritin showed strong inhibition to OAT3 and BCRP, and showed certain inhibition to OATP1 B1, OATP1 B3, OCT2, and P-gp inordinately, but no inhibition to OAT1, OCT1, and BSEP.