辛芍提取物中5种成分在Caco―2细胞中的吸收特性.docVIP

辛芍提取物中5种成分在Caco―2细胞中的吸收特性 　　[摘要] 研究辛芍提取物在Caco-2细胞中的吸收特性。该文建立人源结肠腺癌细胞系Caco-2细胞模型，采用UPLC-MS测定辛芍提取物中芍药内酯苷、没食子酸、咖啡酸、灯盏乙素、灯盏甲素5个活性成分在细胞裂解液中的含量。并用此模型分别研究pH，时间，温度以及药物浓度对辛芍提取物中芍药内酯苷等5种成分的吸收影响，并且考察在P-糖蛋白抑制剂存在与否时5种成分的转运情况，预测这5种成分在Caco-2细胞中的吸收机制。实验结果表明，在37 ℃下，辛芍组方提取物中5种成分的摄取具有一定的时间依赖性；在0.5～12.5 g?L-1，摄取呈浓度依赖性，符合被动扩散过程；在pH 4.0～7.4，咖啡酸、灯盏乙素、灯盏甲素3个成分的细胞吸收量随pH增加而显著降低（P0.05）；在4～37 ℃，咖啡酸的细胞吸收量随温度升高而降低，而其余4个成分随着温度升高而增加；加入P-糖蛋白抑制剂维拉帕米和环孢菌素A，咖啡酸与灯盏乙素的细胞摄取量显著增加（P0.05）。说明辛芍提取物中芍药内酯苷等5种成分的吸收机制可能为被动扩散，且灯盏乙素和咖啡酸可能是P-糖蛋的底物。 　　[关键词]辛芍提取物； Caco-2细胞； 吸收； UPLC-MS； 被动扩散 　　[Abstract] To study the absorption characteristics of Xinshao extracts in Caco-2 cells. In this paper， human colon adenocarcinoma cell line Caco-2 cell model was established， and UPLC-MS method was applied to determinate the contents of five components of Xinshao extracts（albiflorin， gallic acid， caffeic acid， scutellarin and apigenin-7-O-glucronide） in cell lysates. This model was also used to study the effect of different drug concentrations， pH， time and temperature on the absorption of five components， investigate the transport of the five components of Xinshao extracts under the conditions with or without P-glycoprotein inhibitors， and predict the absorption mechanism of these five components in Caco-2 cells. The experimental results showed that the absorption of five components of Xinshao extracts in Caco-2 cells was time-dependent at 37 ℃， and concentration-dependent in the range of 0.5-12.5 g?L-1， with a passive diffusion mechanism. At the pH of 4-7.4， the absorption of caffeic acid， scutellarin and apigenin-7-O-glucronide was significantly declined with the increase of pH（P0.05）. At the temperature of 4 to 37 ℃， the absorption of caffeic acid was declined with the increase of temperature， while the absorption of other four components was increased with the increase of temperature. Compared with the control group， caffeic acid and scutellarin cell absorption was significantly higher（P0.05） after treatment with P-g