[Absorption and transport of 6 coumarins isolated from the roots of Angelica pubescens f. biserrata in human Caco-2 cell monolayer model].
[Absorption and transport of 6 coumarins isolated from the roots of Angelica pubescens f. biserrata in human Caco-2 cell monolayer model].
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DOI：
10.3736/jcim20080413
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发表时间：
2008-04-01
期刊：
影响因子：
--
通讯作者：
Wang, Ying
中科院分区：
文献类型：
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作者：
Yang, Xiu-wei;Guo, Qing-mei;Wang, Ying
目的：研究了从毛当归（Angelica pubescens f.）根中分离得到的6种香豆素类化合物（伞形酮、蛇床子素、Columbianadin、醋酸Columbianadin、Angelol-A和Angelol-B）的吸收和跨上皮转运。方法：采用体外培养的人结肠癌细胞系Caco-2细胞单层模型，研究6种香豆素类化合物在Caco-2细胞中从顶侧（AP）向基底侧（BL）和从BL向AP侧的吸收转运。采用反相高效液相色谱-紫外检测器联用技术对6种香豆素类化合物进行了测定。计算转运参数和表观渗透系数（P（app）），并与普萘洛尔作为高渗透性对照品和阿替洛尔作为低渗透性对照品进行比较。以碘乙酰胺为参比标准品，以MK 571为MRP 2抑制剂，研究当归醇B的转运机制。结果：6种香豆素类化合物的吸收转运均以被动扩散为主。伞形酮、蛇床子素、Columbianadin、醋酸Columbianadin、Angelol-A和Angelol-B从AP侧到BL侧的P（app）值分别为（2.679+/-0.263）x 10（-5），（1.306+/-0.324）x 10（-5），（0.595+/-0.086）x 10（-6），（2.930+/-0.410）x 10（-6），（1.532+/-0.444）x 10（-5）和（1.413+/-0.243）× 10（-5）cm/s，从BL到AP侧为（3.381+/-0.410）x 10（-5），（0.898+/-0.134）× 10（-5）、（0.510+/-0.183）× 10（-6）、（0.222+/-0.025）× 10（-6）、（1.203+/-0.280）× 10（-5）和（0.754+/-0.092）× 10（-5）cm/s。在本试验中，普萘洛尔的P（app）值为2.18 x 10（-5）cm/s，阿替洛尔的P（app）值为2.77 x 10（-7）cm/s。6种香豆素类化合物中，伞形酮、蛇床子素、当归醇A和当归醇B从AP侧到BL侧的P（app）值与普萘洛尔相同，而Columbianadin和醋酸Columbianadin介于普萘洛尔和阿替洛尔之间。当用EBSS代替HBSS，分别用碘乙酰胺或MK-591时，当归醇-B的P（app）与对照组相比无统计学差异。平均回收率中伞形花内酯为（83.31 ± 3.52）%，当归醇A为（77.39 ± 7.38）%，蛇床子素、Columbianadin和Angelol-B在50%~ 65%之间，醋酸Columbianadin低于10%。结论：伞形花内酯、蛇床子素、Columbianadin、醋酸Columbianadin、Angelol-A和Angelol-B在Caco-2细胞中的吸收和转运均以被动扩散为主。伞形酮、蛇床子素、当归醇-A和当归醇-B估计为高度吸收的化合物，Columbianadin和Columbianadenacetate估计为中度吸收的化合物。在Caco-2细胞中，骨化三醇和Columbianadin似乎积聚，并且醋酸Columbianadin可能被代谢。当归醇-B的吸收和转运不受pH的变化和碘乙酰胺或MK 571的存在的影响。
OBJECTIVE: To study the absorption and transepithelial transport of six coumarins (umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B, isolated from the roots of Angelica pubescens f. biserrata) in the human Caco-2 cell monolayer model.METHODS: The in vitro cultured human colon carcinoma cell line, Caco-2 cell monolayer model, was applied to study the absorption and transport of the six coumarins from apical (AP) to basolateral (BL) side and from BL to AP side. The six coumarins were measured by reversed-phase high-performance liquid chromatography (HPLC) coupled with ultraviolet absorption detector. Transport parameters and apparent permeability coefficients (P(app)) were calculated and compared with those of propranolol as a control substance of high permeability and atenolol as a control substance of poor permeability. The transport mechanism of angelol-B was assayed by using iodoacetamide as a reference standard to inhibit ATP-dependent transport and MK571 as a well-known inhibitor of MRP2.RESULTS: The absorption and transport of six coumarins were passive diffusion as the dominating process. The P(app) values of umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B from AP to BL side were (2.679+/-0.263) x 10(-5), (1.306+/-0.324) x 10(-5), (0.595+/-0.086) x 10(-6), (2.930+/-0.410) x 10(-6), (1.532+/-0.444) x 10(-5) and (1.413+/-0.243) x 10(-5) cm/s, and from BL to AP side were (3.381+/-0.410) x 10(-5), (0.898+/-0.134) x 10(-5), (0.510+/-0.183) x 10(-6), (0.222+/-0.025) x 10(-6), (1.203+/-0.280) x 10(-5) and (0.754+/-0.092) x 10(-5) cm/s, respectively. In this assay, the P(app) value of propranolol was 2.18 x 10(-5) cm/s and the P(app) value of atenolol was 2.77 x 10(-7) cm/s. Among the 6 coumarins, the P(app) values of umbelliferone, osthole, angelol-A and angelol-B from AP to BL side were identical with that of propranolol, and columbianadin and columbianetin acetate lied between propranolol and atenolol. When replaced the HBSS with EBSS, and iodoacetamide or MK-591 were used in the experiment, the P(app) of angelol-B had no statistical difference as compared with the control group. In the mean total recoveries, umbelliferone was (83.31+/-3.52)%, angelol-A was (77.39+/-7.38)%, osthole, columbianadin and angelol-B were between 50% to 65%, and columbianetin acetate was lower than 10%. The accumulation rates of osthole and columbianadin in the Caco-2 cells were (36.15+/-5.87)% and (53.90+/-4.39)%, respectively.CONCLUSION: The absorption and transport of umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B are passive diffusion as the dominating process in Caco-2 cell monolayer model. Umbelliferone, osthole, angelol-A and angelol-B are estimated to be highly absorbed compounds, and columbianadin and columbianetin acetate are estimated to be moderately absorbed compounds. In the Caco-2 cells, osthol and columbianadin appear to accumulate, and columbianetin acetate may be metabolized. The absorption and transport of angelol-B are not influenced by the change of pH and the presence of iodoacetamide or MK571.