目的 研究小檗堿對有機陰離子轉(zhuǎn)運體（OATs）的抑制作用及其雙向跨膜轉(zhuǎn)運情況。方法 應用由轉(zhuǎn)染試劑Lipo 3 000介導的動物細胞轉(zhuǎn)基因方法、經(jīng)篩選得到人有機陰離子轉(zhuǎn)運體OAT1、OAT2、OAT3、OAT4、OAT7和URAT1高表達單克隆細胞株；以野生型（WT）細胞為對照組，用各轉(zhuǎn)運體放射性同位素標記底物和抑制劑驗證其轉(zhuǎn)運活性；觀察100 μmol/L小檗堿對各轉(zhuǎn)運體的抑制作用，并測定小檗堿對URAT1轉(zhuǎn)運活性的半數(shù)抑制濃度（IC₅₀）。通過Caco-2細胞模型研究小檗堿的雙向跨膜轉(zhuǎn)運情況。結(jié)果 100 μmol/L小檗堿使OAT1、OAT2、OAT3、OAT4、OAT7和URAT1相對轉(zhuǎn)運活性分別下降至（70.48±4.23）%、（69.13±1.28）%、（72.12±3.28）%、（79.77±6.49）%、（69.51±5.99）%、（38.4±2.67）%；小檗堿對URAT1抑制作用的IC₅₀為13.6 μmol/L；在50和100 μmol/L濃度下，小檗堿在頂側(cè)（AP側(cè)）-基底側(cè)（BL側(cè)）方向的跨膜滲透率P_app（A-B）分別為0.28×10^-6 cm/s和0.40×10^-6 cm/s，其相對應的外排率分別為3.18和3.15。結(jié)論 小檗堿對URAT1的抑制作用較強，對OAT1、OAT2、OAT3、OAT4、OAT7抑制作用相對較弱，小檗堿是一些外排蛋白的底物，為預測小檗堿可能發(fā)生的藥物-藥物相互作用、解釋生物利用度低提供了理論依據(jù)。

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.

國家自然科學基金重點項目（81430096）；國家自然科學基金青年科學基金項目（81503154）