目的 基于藥物轉(zhuǎn)運(yùn)體研究元胡止痛方藥效成分延胡索乙素、巴馬汀、原阿片堿、歐前胡素和異歐前胡素跨血腦屏障轉(zhuǎn)運(yùn)機(jī)制。方法 建立hCMEC/D3-U87雙層細(xì)胞模型研究延胡索乙素、巴馬汀、原阿片堿、歐前胡素和異歐前胡素的跨血腦屏障吸收情況；通過(guò)實(shí)時(shí)熒光定量PCR （qRT-PCR）法測(cè)定hCMEC/D3細(xì)胞中相關(guān)轉(zhuǎn)運(yùn)體蛋白多藥耐藥蛋白1（MDR1）、乳腺癌耐藥蛋白（BCRP）、有機(jī)陽(yáng)離子轉(zhuǎn)運(yùn)體1（OCT1）、OCT2、OCT3、有機(jī)陰離子轉(zhuǎn)運(yùn)體1（OCTN1）、OCTN2、OATP1A2、OATP2B1的mRNA表達(dá)水平；使用6株過(guò)表達(dá)人轉(zhuǎn)運(yùn)體的細(xì)胞株（S2-OCT1、S2-OCT3、S2-OCTN1、S2-OCTN2、HEK293-OATP1A2、HEK293-OATP2B1）進(jìn)行攝取實(shí)驗(yàn)，判斷各藥物成分是否為轉(zhuǎn)運(yùn)體蛋白的底物。結(jié)果 各成分均可以一定程度地跨越血腦屏障，延胡索乙素、歐前胡素、異歐前胡素具有更好的跨血腦屏障特性，巴馬汀和原阿片堿的跨血腦屏障能力較弱。外排型轉(zhuǎn)運(yùn)蛋白MDR1、BCRP和攝入型轉(zhuǎn)運(yùn)蛋白OCTN1、OATP2B1、OATP1A2在hCMECD3細(xì)胞中有相對(duì)高的mRNA表達(dá)水平。延胡索乙素是OCT1、OCTN1和OATP1A2的底物；巴馬汀是OCT1、OCT3、OCTN1、OCTN2和OATP2B1的底物；原阿片堿是OCT1、OCTN1和OCTN2的底物；歐前胡素、異歐前胡素是OCT3的底物。結(jié)論 延胡索乙素可能通過(guò)轉(zhuǎn)運(yùn)體OATP1A2、OCT1、OCTN1跨越血腦屏障進(jìn)入腦組織細(xì)胞；巴馬汀可能通過(guò)OATP2B1、OCT1、OCT3、OCTN1、OCTN2傳遞進(jìn)入腦組織細(xì)胞；原阿片堿可能通過(guò)OCT1、OCTN1和OCTN2傳遞進(jìn)入腦組織細(xì)胞；歐前胡素、異歐前胡素可能通過(guò)OCT3傳遞進(jìn)入腦組織細(xì)胞。

Objective To explore the trans-blood-brain barrier (BBB) transport mechanism of different medicinal components of Yuanhu Zhitong Prescription: tetrahydropalmatine, palmatine, fumarine, imperatorin and isoimperatorin based on drug transporter. Methods A bilayer cell model of hCMEC/D3-U87 was developed to study the cross-BBB absorption of different components of the drug. The mRNA expression levels of related transporter proteins multidrug resistance protein 1 (MDR1), breast cancer resistance protein (BCRP), organic cation transporter 1 (OCT1), OCT2, OCT3, organic anion transporter 1 (OCTN1), OCTN2, OATP1A2, OATP2B1 in hCMEC/D3 cells were measured by real-time fluorescence quantitative PCR. Six cell lines overexpressing human transporters (S2-OCT1, S2-OCT3, S2-OCTN1, S2-OCTN2, HEK293-OATP1A2, HEK293-OATP2B1) were used for the uptake test to determine whether the drug components were substrates of transporter protein. Results All components could cross the bloodbrain barrier to a certain extent, tetrahydropalmatine, imperatorin and isoimperatorin had better characteristics of crossing the bloodbrain barrier, while palmatine and protoopiate had weaker ability of crossing the blood-brain barrier. The efflux transporter MDR1, BCRP and the uptake transporter OCTN1, OATP2B1, OATP1A2 had relatively high mRNA expression levels in hCMECD/3 cells. Tetrahydropalmatine was the substrate of OCT1, OCTN1 and OATP1A2. Palmatine was the substrate of OCT1, OCT3, OCTN1, OCTN2 and OATP2B1. Protopine was the substrate of OCT1, OCTN1 and OCTN2. Imperatorin and isoimperatorin were the substrates of OCT3. Conclusion Tetrahydropalmatine enters the BBB via OATP1A2, OCT1, and OCTN1. Palmatine enters the BBB via OATP2B1, OCT1, OCT3, OCTN1, and OCTN2. Protopine enters the BBB via OCT1, OCTN1, and OCTN2.Imperatorin and isoimperatorin enters the BBB via OCT3.

R962.2

釋藥技術(shù)與藥代動(dòng)力學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室（天津藥物研究院）自主研究課題資助（010161002）；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（2019-I2M-5-020）；天津市“項(xiàng)目+團(tuán)隊(duì)”重點(diǎn)培養(yǎng)專項(xiàng)（創(chuàng)新類）（XC202030）