目的 基于药物转运体研究元胡止痛方药效成分延胡索乙素、巴马汀、原阿片碱、欧前胡素和异欧前胡素跨血脑屏障转运机制。方法 建立hCMEC/D3-U87双层细胞模型研究延胡索乙素、巴马汀、原阿片碱、欧前胡素和异欧前胡素的跨血脑屏障吸收情况；通过实时荧光定量PCR （qRT-PCR）法测定hCMEC/D3细胞中相关转运体蛋白多药耐药蛋白1（MDR1）、乳腺癌耐药蛋白（BCRP）、有机阳离子转运体1（OCT1）、OCT2、OCT3、有机阴离子转运体1（OCTN1）、OCTN2、OATP1A2、OATP2B1的mRNA表达水平；使用6株过表达人转运体的细胞株（S2-OCT1、S2-OCT3、S2-OCTN1、S2-OCTN2、HEK293-OATP1A2、HEK293-OATP2B1）进行摄取实验，判断各药物成分是否为转运体蛋白的底物。结果 各成分均可以一定程度地跨越血脑屏障，延胡索乙素、欧前胡素、异欧前胡素具有更好的跨血脑屏障特性，巴马汀和原阿片碱的跨血脑屏障能力较弱。外排型转运蛋白MDR1、BCRP和摄入型转运蛋白OCTN1、OATP2B1、OATP1A2在hCMECD3细胞中有相对高的mRNA表达水平。延胡索乙素是OCT1、OCTN1和OATP1A2的底物；巴马汀是OCT1、OCT3、OCTN1、OCTN2和OATP2B1的底物；原阿片碱是OCT1、OCTN1和OCTN2的底物；欧前胡素、异欧前胡素是OCT3的底物。结论 延胡索乙素可能通过转运体OATP1A2、OCT1、OCTN1跨越血脑屏障进入脑组织细胞；巴马汀可能通过OATP2B1、OCT1、OCT3、OCTN1、OCTN2传递进入脑组织细胞；原阿片碱可能通过OCT1、OCTN1和OCTN2传递进入脑组织细胞；欧前胡素、异欧前胡素可能通过OCT3传递进入脑组织细胞。

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

释药技术与药代动力学国家重点实验室（天津药物研究院）自主研究课题资助（010161002）；中国医学科学院医学与健康科技创新工程项目（2019-I2M-5-020）；天津市“项目+团队”重点培养专项（创新类）（XC202030）