杜仲提取物中4种主要成分在Caco-2细胞的摄取特性研究

Uptake of Eucommia ulmoides extract in Caco-2 cell monolayer

目的研究杜仲提取物中京尼平苷酸、原儿茶酸、松脂醇二葡萄糖苷、松脂醇单葡萄糖苷在Caco-2细胞的摄取特性。方法以Caco-2细胞单层模型研究杜仲提取物的细胞摄取规律,采用UPLC-MS/MS法测定Caco-2细胞中京尼平苷酸、原儿茶酸、松脂醇二葡萄糖苷、松脂醇单葡萄糖苷的浓度,考察时间、pH值、药物浓度、温度及抑制剂对Caco-2细胞摄取杜仲提取物的影响。结果杜仲提取物中京尼平苷酸、原儿茶酸、松脂醇二葡萄糖苷、松脂醇单葡萄糖苷4种成分在Caco-2细胞中的摄取具有一定的时间、浓度依懒性,其摄取表现为被动扩散;在pH 4条件下,杜仲提取物中上述4种成分的细胞摄取量明显高于pH 8;在4℃、25℃、37℃条件下,杜仲提取物在37℃下的摄取量最高;加入维拉帕米、环孢菌素A后,杜仲提取物中原儿茶酸的细胞摄取量有明显变化,其余3个成分没有变化。结论杜仲提取物中京尼平苷酸、原儿茶酸、松脂醇二葡萄糖苷、松脂醇单葡萄糖苷4种成分的细胞摄取机制主要是被动转运;P-糖蛋白参与其原儿茶酸的摄取过程。

Aim To observe the uptake process of geniposidic acid , protocatechuic acid , pinoresinol diglucoside and pnoresinol singleglucoside in Caco-2 cell. Methods The concentration of the four componds in cells was detected by UPLC-MS/MS, and the mechanism of the four componds absorption in cells was explored by studying the time, pH, drug concen-tration and inhibitors on the uptake of Eucommia ul-moides extract in cells. Results Eucommia ulmoides extract in Caco-2 cell uptake was time-dependent. Eu-commia ulmoides extract at 0.2~5 g·L-1 concentra-tion range of uptake increased linearly, consistent with passive diffusion process. The drug uptake amount un-der pH 4.0 was significantly higer than pH 8 . 0 （ P〈0.05 ） . The drug uptake amount was the highest in 37℃. Compared with control group, geniposidic acid, protocatechuic acid, pinoresinol diglucoside and pnoresinol singleglucoside cell uptake was not signifi-cantly higher after additional treatment with verapamil and cyclosporin A, and protocatechuic acid cell uptake was significantly higher after additional treatment with verapamil and cyclosporin A, Conclusion The mech-anism of Eucommia ulmoides extract absorption in cells was mainly through passive transport, and P-glycopro-teins participates in the conveying process of protocate-chuic acid in Caco-2 cells.