目的 研究6种药物对有机阴离子转运多肽OATP1B1及其基因多态性A388G、T521C转运作用的影响。方法 体外培养稳定高表达OATP1B1和OATP1B1基因多态性A388G、T521C的人胚肾细胞（HEK293）株，高表达空白载体（Mock）的HEK293细胞为空白对照，实时荧光定量PCR（qRT-PCR）法检测各转运体细胞中mRNA表达；放射性标记化合物³Hestrone sulfate作为转运底物、利福平作为阳性抑制剂验证各高表达细胞的转运活性；测定30 μmol·L^-1的达比加群、辛伐他汀、替格瑞洛、卡培他滨、多西他赛、依那普利对各细胞³H-estrone sulfate摄入活性的抑制作用，并依据抑制试验结果，进一步测定辛伐他汀、替格瑞洛、多西他赛对转运体细胞的半数抑制浓度（IC₅₀）。结果 HEK293细胞内导入的各种转运体基因都呈现良好的复制表达；OATP1B1、OATP1B1/A388G、OATP1B1/T521C对底物³H-estrone sulfate（5 μmol·L^-1）的转运活性分别为Mock细胞的39、49和48倍，30 μmol·L^-1利福平添加后，可将细胞的转运活性抑制到50%以下；辛伐他汀、替格瑞洛、多西他赛对OATP1B1的抑制作用较强，30 μmol·L^-1给药的转运活性分别为对照组的（40.09±1.95）%、（33.82±0.61）%、（45.08±0.22）%；辛伐他汀对OATP1B1、OATP1B1/A388G、OATP1B1/T521C的IC₅₀分别为14.2、＞100、＞100 μmol·L^-1，替格瑞洛的IC₅₀分别为19.1、68.4、＞100 μmol·L^-1，多西他赛的IC₅₀分别为17.6、22.9、19.3 μmol·L^-1。结论 OATP1B1基因多态性在一定程度上改变了抑制剂对转运体活性的影响程度。

Objective Study the transport effect of six drugs on the organic anion transporting polypeptide OATP1B1 and its gene polymorphisms A388G and T521C. Methods Human embryonic kidney cell (HEK293) lines stably and highly expressing OATP1B1 and OATP1B1 gene polymorphisms A388G and T521C were cultured in vitro, HEK293 cells with high expression of blank vector (Mock) were used as blank control. The Real time fluorescent quantitative PCR (qRT-PCR) method was used to detect mRNA expression in each transporter cell. The radiolabelled compound ³H-estrone sulfate was used as a transport substrate to verify the transport activity of each highly expressed cell. The inhibitory effects of 30 μmol·L^-1 dabigatran, simvastatin, Tegretol, capecitabine, docetaxel and enalapril on the transport activity of ³H-estrone sulfate in each cell were determined, and the median inhibitory concentration (IC₅₀) of simvastatin, ticagrelor and docetaxel on transporter cells was further determined based on the results of the inhibition test. Results Various transporter genes introduced into HEK293 cells showed good replication and expression. The transport activity of OATP1B1, OATP1B1/A388G and OATP1B1/T521C to substrate ³H-estrone sulfate (5 μmol·L^-1) was 39, 49 and 48 times that of Mock cells, respectively. Rifampicin of 30 μmol·L^-1 could inhibit the cell transport activity to less than 50%. The inhibition of OATP1B1 by 30 μmol·L^-1 simvastatin, tegretol, and docetaxel was stronger, were (40.09±1.95)%, (33.82±0.61)%, and (45.08±0.22)% of control group, respectively. The IC₅₀ of simvastatin on OATP1B1, OATP1B1/A388G, and OATP1B1/T521C were 14.2, >100, >100 μmol·L^-1, IC₅₀ of tegretol were 19.1, 68.4, >100 μmol · L^-1 and IC₅₀ of docetaxel were 17.6, 22.9, and 19.3 μmol·L^-1, respectively. Conclusion The polymorphism of OATP1B1 gene changed the effect of inhibitor on transporter activity to some extent.

R969.2

中国医学科学院医学与健康科技创新工程项目资助（2019-I2M-5-020）