目的 研究6種藥物對(duì)有機(jī)陰離子轉(zhuǎn)運(yùn)多肽OATP1B1及其基因多態(tài)性A388G、T521C轉(zhuǎn)運(yùn)作用的影響。方法 體外培養(yǎng)穩(wěn)定高表達(dá)OATP1B1和OATP1B1基因多態(tài)性A388G、T521C的人胚腎細(xì)胞（HEK293）株，高表達(dá)空白載體（Mock）的HEK293細(xì)胞為空白對(duì)照，實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測(cè)各轉(zhuǎn)運(yùn)體細(xì)胞中mRNA表達(dá)；放射性標(biāo)記化合物³Hestrone sulfate作為轉(zhuǎn)運(yùn)底物、利福平作為陽性抑制劑驗(yàn)證各高表達(dá)細(xì)胞的轉(zhuǎn)運(yùn)活性；測(cè)定30 μmol·L^-1的達(dá)比加群、辛伐他汀、替格瑞洛、卡培他濱、多西他賽、依那普利對(duì)各細(xì)胞³H-estrone sulfate攝入活性的抑制作用，并依據(jù)抑制試驗(yàn)結(jié)果，進(jìn)一步測(cè)定辛伐他汀、替格瑞洛、多西他賽對(duì)轉(zhuǎn)運(yùn)體細(xì)胞的半數(shù)抑制濃度（IC₅₀）。結(jié)果 HEK293細(xì)胞內(nèi)導(dǎo)入的各種轉(zhuǎn)運(yùn)體基因都呈現(xiàn)良好的復(fù)制表達(dá)；OATP1B1、OATP1B1/A388G、OATP1B1/T521C對(duì)底物³H-estrone sulfate（5 μmol·L^-1）的轉(zhuǎn)運(yùn)活性分別為Mock細(xì)胞的39、49和48倍，30 μmol·L^-1利福平添加后，可將細(xì)胞的轉(zhuǎn)運(yùn)活性抑制到50%以下；辛伐他汀、替格瑞洛、多西他賽對(duì)OATP1B1的抑制作用較強(qiáng)，30 μmol·L^-1給藥的轉(zhuǎn)運(yùn)活性分別為對(duì)照組的（40.09±1.95）%、（33.82±0.61）%、（45.08±0.22）%；辛伐他汀對(duì)OATP1B1、OATP1B1/A388G、OATP1B1/T521C的IC₅₀分別為14.2、＞100、＞100 μmol·L^-1，替格瑞洛的IC₅₀分別為19.1、68.4、＞100 μmol·L^-1，多西他賽的IC₅₀分別為17.6、22.9、19.3 μmol·L^-1。結(jié)論 OATP1B1基因多態(tài)性在一定程度上改變了抑制劑對(duì)轉(zhuǎn)運(yùn)體活性的影響程度。

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

中國(guó)醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程項(xiàng)目資助（2019-I2M-5-020）