MDCK-MDR1细胞药物体外吸收模型的建立及验证  

ESTABLISHMENT AND VALIDATION OF A DRUG IN VITRO ABSORPTION MODEL OF MDCK-MDR1 CELLS

作　　者：韩帅 张洁[2] 金婷婷 闫茹冰 褚海辰 梁永新[1,3] HAN Shuai;ZHANG Jie;JIN Tingting;YAN Rubing;CHU Haichen;LIANG Yongxin(Faculty of Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学医学部,山东青岛266071 [2]青岛大学附属医院麻醉科 [3]青岛大学附属妇女儿童医院麻醉科

出　　处：《精准医学杂志》2023年第2期145-148,153,共5页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81873729)。

摘　　要：目的构建MDCK-MDR1细胞体外吸收模型并进行验证,以用于口服药物吸收和转运机制的研究。方法将不同浓度组(1.0×10^(8)/L的L组、2.5×10^(8)/L的M组、5.0×10^(8)/L的H组)MDCK-MDR1细胞接种于24孔Transwell培养板上,培养1~7 d,通过吸光度值绘制MDCK-MDR1细胞生长曲线,观察不同培养时间点细胞的形态,测定不同时间点的跨膜电阻(TEER)值,确定形成单层膜结构的最佳细胞接种浓度和培养时间。通过荧光黄转运实验对不同浓度和时间点形成的单细胞膜结构进行验证。结果L组、M组和H组分别在接种后第5、3、1天形成单层膜结构,分别在第5、4、3天时吸光度值达到峰值。L组在接种第5天时TEER值达到300Ω·cm^(2),第5~7天趋于稳定,故确定形成单层膜结构的最佳细胞接种浓度为1.0×10^(8)/L,最佳培养时间为5 d。荧光黄转运实验验证显示,该单层膜结构的荧光黄表观渗透系数为4.27×10^(-7)cm/s,低于通透性试验规定的5.0×10^(-7)cm/s。结论本研究构建的MDCK-MDR1细胞模型单层膜结构完整性和通透性均通过验证,可作为模拟口服药物吸收和转运机制研究的体外模型。Objective To establish and validate a drug in vitro absorption model of MDCK-MDR1 cells for research on the mechanism of oral drug absorption and transport.Methods MDCK-MDR1 cells with different concentrations(group L with 1.0×10^(8)/L,group M with 2.5×10^(8)/L,and group H with 5.0×10^(8)/L)were inoculated onto a 24-well Transwell plate and cultured for 1-7 d.Optical density(OD)was measured to plot the growth curve of MDCK-MDR1 cells,and cell morphology was observed at different time points of culture.Transepithelial electrical resistance(TEER)was measured at different time points to determine the optimal cell inoculation concentration and culture time for the formation of a monolayer structure.Lucifer yellow transfer assay was used to validate the monolayer structure formed at different concentrations and time points.Results The groups L,M,and H showed the formation of the monolayer structure on days 5,3,and 1,respectively,after inoculation,and OD reached the peak value on days 5,4,and 3,respectively,For group L,TEER reached 300Ω·cm^(2)on day 5 of inoculation and tended to be stable on days 5-7.Therefore,1.0×10^(8)/L was selected as the optimal cell inoculation concentration,with an optimal culture time of 5 d.Lucifer yellow transfer assay showed that the monolayer structure had a Lucifer yellow apparent permeability coefficient of 4.27×10^(-7)cm/s,which was lower than 5.0×10^(-7)cm/s determined by the permeability test.Conclusion The MDCK-MDR1 cell model established in this study has validated monolayer structural integrity and permeability and can be used as an in vitro model to simulate the absorption and transport mechanisms of oral drugs.

关 键 词：MDCK-MDR1细胞 电阻抗 细胞膜通透性 模型 生物学 生物转运 肠吸收 体外培养技术 

分 类 号：R349.92[医药卫生—基础医学] R329.27