用于中药复方物质基础研究的Caco-2细胞胶原包被6孔转运板体系的构建  

Establishment of Caco-2 cell monolayer model with collagen coating 6-well plates for study of traditional Chinese medicine prescription

作　　者：杨雁芳[1] 吴妮[1] 杨秀伟[1] 

机构地区：[1]北京大学药学院天然药物学系 天然药物及仿生药物国家重点实验室,北京100191

出　　处：《中国中药杂志》2014年第3期515-519,共5页China Journal of Chinese Materia Medica

基　　金：国家“重大新药创制”科技重大专项(2009ZX09502-006);国家科技支撑计划项目(2011BAI07B08)

摘　　要：6孔转运板胶原包被后进行Caco-2单层细胞培养,测定不同的胶原种类、鼠尾胶原铺胶浓度和铺胶时间下Caco-2细胞单层的跨膜电阻以及阳性药物普萘洛尔和阿替洛尔的被动扩散表观渗透系数,发现不同胶原包被条件对Caco-2细胞单层的完整性和转运能力无明显影响。因此2μg·cm-2鼠尾胶原铺胶1 h即可构建稳定的Caco-2细胞培养的胶原包被6孔转运板体系,为研究中药复方物质基础的吸收提供体外研究技术方法。Caco-2 cell monolayer model is widely utilized in drug absorption study and 12-well transwellTM plates were com- monly used to study the absorption of different kinds of natural products. To establish a stable method for the study of traditional Chi- nese medicine prescription, 6-well plates were chosen because of the larger well volumes than 12-well plates. To study the impacts of collagen kinds, coating density as well as coating time on the cell culture, the transepithelial electrical resistance of Caco-2 cell monolayers grown on different collagen coating transwells was determined, and the permeations of propranolol and atenolol as standard markers were detected with HPLC. The results showed that the kinds of collagen, the different coating densities and coating time of rat tail collagen had no significant influences on the Caco-2 cell monolayer integrality and absorption capacity. 6-well plates coated with 2 ug . cm-2 rat tail collagen for 1 hour were enough reliable and suitable for the study of traditional Chinese medicine prescription in vitro.

关 键 词：CACO-2细胞单层 6孔板 胶原 跨膜电阻 表观渗透系数 

分 类 号：R284[医药卫生—中药学]