中药化学成分肠吸收研究中Caco-2细胞模型和标准操作规程的建立

目的:建立人结肠腺癌细胞系(human colon adenocarcinoma cell line,Caco-2)细胞模型和标准操作规程,用于研究和评价中药化学成分的肠吸收。方法:通过电镜扫描和倒置显微镜观察Caco-2细胞单层细胞形态学特点,并通过测定跨Caco-2细胞单层细胞膜电阻、碱性磷酸酶活性以及易吸收阳性对照药普萘洛尔和难吸收阳性对照药阿替洛尔的转运特性等指标对Caco-2细胞模型进行评价。结果:建立的Caco-2细胞模型完整性、紧密性和通透性等良好,各项指标的测定值与文献值一致。结论:建立的Caco-2细胞单昙模型符合各项指标的要求,可用于研究口服中药化学成分的肠吸收及其转运的吸收机制。

Establishment of Caco-2 cell monolayer model and standard operation procedure for assessing intestinal absorption of chemical components of traditional Chinese medicine

Objective:To establish Caco-2(a human colon adenocarcinoma cell line)cell monolayer model and the standard operation procedure for studying and assessing intestinal absorption of chemical components of traditional Chinese medicine. Methods.Caco-2 cell monolayer model was established and evaluated by morphology feature using scanning electron microscope,inverted microscope and transepithelial electrical resistance(TEER)assay. Additionally.the model was further tested for the activity of alkaline phosphatase and the apparent permeability(P_(app))of standard compounds,i.e.propranolol and atenolol,which were the control substances for high and poor transcellular transport marker,respectively. Results.The integrality of cell monolayer,cell differentiation(reflected by expression of alkaline phosphatase and cell monolayer morphology),and the P_(app)value of standard compounds in the established Caco-2 cell model were satisfactory.All parameters tested were in good agreement with those reported in the literature. Conclusion:The established Caco-2 cell model can be used to study the intestinal absorption of orally administrated chemical components of traditional Chinese medicine and their absorption mechanism.