Caco-2细胞中药物转运蛋白的mRNA表达及活力评价  

Expression of drug transporters mRNA in Caco-2 cells and its viability evaluation

作　　者：慈小燕[1] 王泽 李亚卓[1] 谷元[1] 武卫党[1] 曾勇[1] 伊秀林[1] 司端运[1] CI Xiaoyan;WANG Ze;LI Yazhuo;GU Yuan;WU Weidang;ZENG Yong;YI Xiulin;SI Duanyun(State Key Laboratory of Drug Delivery Technology and Pharmacokinetics,Tianjin Institute of Pharmaceutical Research New Drug Evaluation Co.Ltd.,Tianjin 300193,China;Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China)

机构地区：[1]天津药物研究院新药评价有限公司释药技术与药代国家重点实验室,天津300193 [2]天津中医药大学,天津300193

出　　处：《药物评价研究》2018年第6期980-985,共6页Drug Evaluation Research

基　　金：国家自然科学基金青年科学基金项目(81503154);国家重点研发计划应用系统生物学结合PK-PD研究乌头汤药性配伍理论(2016YFE0121400)

摘　　要：目的系统研究Caco-2细胞中各药物转运蛋白的m RNA表达水平及转运活力,对比其与人正常肠道中药物转运蛋白表达的差异。方法实时荧光定量PCR(q RT-PCR)方法测定Caco-2细胞中人肠道相关转运蛋白MDR1、BCRP、MRP2、OATP1A2、OATP2B1和PEPT1的表达水平;将Caco-2细胞接种于Transwell板内培养21 d并给予不同药物转运蛋白的底物及抑制剂,评价Caco-2细胞中相关转运蛋白的转运活性。结果 q RT-PCR结果表明,药物转运蛋白MDR1、MRP2、BCRP和OATP2B1在Caco-2细胞中均有相对高的表达,表达量的顺序为:MDR1>MRP2>OATP2B1>BCRP,在正常人肠道表达量顺序为BCRP>MDR1>MRP2>OATP2B1;转运蛋白活力评价表明,各药物转运蛋白的活力测试结果均为阳性,验证了基因的表达水平结果。结论 Caco-2细胞中表达正常人体肠道表达的部分药物转运蛋白(MDR1、MRP2、BCRP和OATP2B1),表达水平与正常人体肠道中大致相当,但也存在一定差异。Objective To study the m RNA expression and transport activity of drug transporters in Caco-2 cells, and compare the differences of the expression of drug transporters between Caco-2 cells and human normal intestinal. Method Establishing a method for the determination of m RNA expression of drug transporter by q RT-PCR method to determine the expression levels of human intestinal related transport proteins MDR1, BCRP, MRP2, OATP1 A2, OATP2 B1, and PEPT1 in Caco-2 cells; Caco-2 was then seeded in Transwell plates for 21 days and administered with different drug transporter substrates and inhibitors to assess the transporter activity of the relevant transporter in Caco-2 cells. Results The q RT-PCR results indicated that MDR1, MRP2, BCRP, and OATP2 B1 were relatively higher expressed in Caco-2 cells. The relative expression levels of various drug transporters were as follows： MDR1 〉MRP2〉 OATP2B1〉 BCRP, the expression level of intestinal tract in normal subjects was BCRP〉 MDR1〉 MRP2〉 OATP2 B1. Drug transporter activity assessment showed that all drug transporter activity test results were positive, validating the expression level of the previous drug transporter gene. Conclusions The expression levels of many drug transporters（MDR1, MRP2, BCRP and OATP2 B1） originated from normal human intestinal tract expressed in Caco-2 cells were roughly equivalent to those expressed in the normal human intestinal tract, but there were also some differences between them.

关 键 词：CACO-2细胞 肠道渗透性 药物转运蛋白 实时荧光定量PCR 表观渗透系数 

分 类 号：R962.2[医药卫生—药理学]