目的 研究小檗碱对有机阴离子转运体（OATs）的抑制作用及其双向跨膜转运情况。方法 应用由转染试剂Lipo 3 000介导的动物细胞转基因方法、经筛选得到人有机阴离子转运体OAT1、OAT2、OAT3、OAT4、OAT7和URAT1高表达单克隆细胞株；以野生型（WT）细胞为对照组，用各转运体放射性同位素标记底物和抑制剂验证其转运活性；观察100 μmol/L小檗碱对各转运体的抑制作用，并测定小檗碱对URAT1转运活性的半数抑制浓度（IC₅₀）。通过Caco-2细胞模型研究小檗碱的双向跨膜转运情况。结果 100 μmol/L小檗碱使OAT1、OAT2、OAT3、OAT4、OAT7和URAT1相对转运活性分别下降至（70.48±4.23）%、（69.13±1.28）%、（72.12±3.28）%、（79.77±6.49）%、（69.51±5.99）%、（38.4±2.67）%；小檗碱对URAT1抑制作用的IC₅₀为13.6 μmol/L；在50和100 μmol/L浓度下，小檗碱在顶侧（AP侧）-基底侧（BL侧）方向的跨膜渗透率P_app（A-B）分别为0.28×10^-6 cm/s和0.40×10^-6 cm/s，其相对应的外排率分别为3.18和3.15。结论 小檗碱对URAT1的抑制作用较强，对OAT1、OAT2、OAT3、OAT4、OAT7抑制作用相对较弱，小檗碱是一些外排蛋白的底物，为预测小檗碱可能发生的药物-药物相互作用、解释生物利用度低提供了理论依据。

Objective To study the inhibition of berberine on organ anion transporters(OATs) and its bidirectional trans-membrane transport. Method The transgene cell lines of the organ anion transporters including OAT1, OAT2, OAT3, OAT4, OAT7, and URAT1 were constructed and selected by animal cell transgenic method mediated by transporter Lipo 3000.Wild type (WT) cells were used as control group, and activity of OATs was verified by adding their radiolabeled substrates and inhibitors. The inhibition of 100 μmol/Lberberine on the transporters was investigated in vitro. The IC₅₀ of berberine on URAT1 was also determined.The bidirectional transport of berberine was studied through the Caco-2 model. Result The results showed that 100 μmol/L berberine inhibited the activity of OAT1, OAT2, OAT3, OAT4, OAT7 and URAT1 to (70.48±4.23)%, (69.13±1.28)%, (72.12±3.28)%, (79.77±6.49)%, (69.51±5.99)% and (38.4±2.67)% respectively, the IC₅₀ of berberine to URAT1 was 13.19 μmol/L, the P_{app (A-B)} of 50 μmol/L and 100 μmol/L berberine were separately 0.28×10^-6 and 0.40 ×10^-6 cm/s, and the efflux rates were separately 3.18 and 3.15. Conclusion Berberine shows a stronger inhibition to URAT1 compared to OAT1, OAT2, OAT3, OAT4 and OAT7. Berberine may be the substrate of some efflux transporters.This study provides theoretical basis for explaining the low bioavailability of berberine and forecasting the possible drug-drug interaction.

国家自然科学基金重点项目（81430096）；国家自然科学基金青年科学基金项目（81503154）