Quantitative Comparison of Bile Acid Distribution and Intestinal Transport from Native Cow-bezoar and Artificial and in vitro Cultured Substitutes using Caco-2 Cell Monolayer Model

天然牛黄、体外培育牛黄和人工牛黄中胆汁酸类成分及其在人肠源Caco-2细胞模型中吸收转运的对比研究（英文）

Objective This study was conducted to examine the absorption and translocation of conjugated bile acids(BAs)in Calculus bovis and its substitutes to detect differences in these materials.Methods A Caco-2 monolayer cell model was used to compare the apparent permeability coefficient(Papp)value and efflux ratio(ER)of BAs in natural cow-bezoar(NCB),artificial cow-bezoar(ACB),and in vitro cultured cow-bezoar(Ivt-CCB).Papp and ER values were determined by liquid chromatography-mass spectrometry.Samples were separated on an analytical column.Results The distribution of BAs in NCB was significantly different from that in ACB and Ivt-CCB.The percentages of conjugated BAs were significantly higher in NCB than in the two substitutes.The distribution differences of conjugated and unconjugated BAs can be used to distinguish costly NCB from relatively inexpensive substitutes.Conclusion The transport characteristics of BAs in Ivt-CCB were more consistent with NCB than with ACB,even when the proportions of BAs in Ivt-CCB were closer to those of ACB.

目的对天然牛黄、体外培育牛黄及人工牛黄中的多种胆汁酸在体内吸收及转运进行研究。方法建立Caco-2细胞单层膜转运模型,收集天然牛黄、体外培育牛黄和人工牛黄转运前后的样品。采用Acquity UPLC HSS T3色谱柱(2.1 mm×100 mm; 1.8μm),以10 mmol/L醋酸铵为流动相A,以乙腈-甲醇(3:1)为流动相B,梯度洗脱(0～1 min,2%B→15%B;4～10 min,40%B→60%B;14～22 min,95%B;流速0.3 mL/min),采用电喷雾离子源进行负离子模式扫描,对牛黄及代用品中甘氨胆酸、牛磺胆酸、去氧胆酸、胆酸等胆汁酸类成分在Caco-2细胞转运模型上的表观通透系数(Papp)和外排率(ER)进行测定。结果人工牛黄和体外培育牛黄中大多数胆汁酸类成分的双向透过率均低于天然牛黄;天然牛黄和人工牛黄中胆酸、鹅去氧胆酸、牛磺去氧胆酸和甘氨去氧胆酸的分泌外排率存在显著差异,但体外培育牛黄与天然牛黄没有显著差异。结论牛黄、体外培育牛黄中胆汁酸类成分在Caco-2细胞上的转运机制可能较为相似。