Caco-2细胞单层模型中水飞蓟宾吸收机制研究

Absorption mechanism of silybin in human intestinal Caco-2 cells

通过建立Caco-2细胞单层模型考察了不同浓度水平的水飞蓟宾和水飞蓟宾葡甲胺的双向跨膜通透性并研究其吸收机制。开发HBSS缓冲液中水飞蓟宾、普萘洛尔和阿替洛尔的LC/MS/MS检测方法,检测实验样品的浓度,并计算表观渗透系数(P_(app))。当Caco-2细胞培养到第21天时,对所建立的Caco-2细胞模型进行验证:跨膜电阻值(TEER)大于350Ω·cm2,荧光黄的P_(app)远远小于1×10^(-7)cm/s,阳性对照药阿替洛尔和普萘洛尔的P_(app)与文献中报道的相近,表明Caco-2细胞单层模型建立成功。3个浓度水平的水飞蓟宾(5,20,50μg/m L)P_(app(AP-BL))均大于2×10^(-6)cm/s,证明水飞蓟宾的通透性良好。P_(app)(BL-AP)与P_(app(AP-BL))的比值大于2,说明在水飞蓟宾吸收过程中有外排转运体的参与。水飞蓟宾葡甲胺的P_(app)与水飞蓟宾的基本一致,成盐对于水飞蓟宾的跨膜通透性没有改变。结果表明,水飞蓟宾为跨膜通透性良好药物,其溶解度低,在生物药剂学分类系统(BCS)分类上为Ⅱ类。水飞蓟宾在胃肠道中的释放是其吸收过程的重要影响因素。

The aim of this paper was to investigate the absorption mechanism of silybin(SLB)in Caco-2 cells.Concentrations of samples in the study were determined by developing LC/MS/MS method of SLB,propranolol and atenolol in HBSS buffer to calculate apparent permeability coefficient(P app).When Caco-2 cells were cultured to the 21st day,the TEER were above 350Ω·cm 2.The P app of Lucifer yellow was far less than 1×10-7 cm/s.As the positive control drugs,The P app of atenolol and propranolol were similar to those reported in the literature,indicating that the Caco-2 monolayer model was successfully established in this experiment.The P app(AP-BL)of SLB at 5,20,and 50μg/mL were all more than 2×10-6 cm/s,which showed that SLB was a moderately permeable drug.The efflux ratio was greater than 2 indicating the efflux transporter may be involved in the absorption process of SLB.The P app of silybin-N-meglumine was similar to that of SLB,suggesting that salt formation did not alter the membrane permeability of SLB.In conclusion,the membrane permeability of SLB is good,and its solubility is low.SLB is a BCS class 2 drug.The release of SLB in the gastrointestinal tract is an important factor in its absorption process.