Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (4): 392-398.doi: 10.3736/jcim20080413

• Original Experimental Research • Previous Articles     Next Articles

Absorption and transport of 6 coumarins isolated from the roots of Angelica pubescens f. biserrata in human Caco-2 cell monolayer model

Xiu-wei Yang1(), Qing-mei Guo2, Ying Wang1   

  1. 1. Department of Natural Medicines,State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,Peking University, Beijing 100083, China
    2. College of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province 250014, China
  • Received:2007-08-02 Online:2008-04-20 Published:2008-04-15
  • Contact: YANG Xiu-wei E-mail:xwyang@bjmu.edu.cn

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 (Papp) 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 Papp values of umbelliferone, osthole, columbianadin, columbianetin acetate, angelol-A and angelol-B from AP to BL side were (2.679±0.263)×10 -5, (1.306±0.324)×10 -5, (0.595±0.086)×10 -6, (2.930±0.410)×10 -6, (1.532±0.444)×10 -5 and (1.413±0.243)×10 -5 cm/s, and from BL to AP side were (3.381±0.410)×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 and (0.754±0.092)×10 -5 cm/s, respectively. In this assay, the Papp value of propranolol was 2.18×10 -5 cm/s and the Papp value of atenolol was 2.77×10 -7 cm/s. Among the 6 coumarins, the Papp 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 Papp 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 metabolited. The absorption and transport of angelol-B are not influenced by the change of pH and the presence of iodoacetamide or MK571.

Key words: Caco-2 cell, coumarins, intestinal absorption, apparent permeability coefficient

CLC Number: 

  • R285.5

Figure 1

Chemical structures of 6 coumarins 1: Umbelliferone; 2: Osthole; 3: Columbianadin; 4: Columbianetin acetate; 5: Angelol-A; 6: Angelol-B."

Figure 2

Speed-rate of 6 coumarins transported from AP side to BL side in the Caco-2 cell monolayer model 1: Umbelliferone; 2: Osthole; 3: Columbianadin; 4: Columbianetin acetate; 5: Angelol-A; 6: Angelol-B. Coumarins were incubated with Caco-2 cells for 90 min. The values presented M±SD (n=3)."

Figure 3

Speed-rate of 6 coumarins transported from BL side to AP side in the Caco-2 cell monolayer model 1: Umbelliferone; 2: Osthole; 3: Columbianadin; 4: Columbianetin acetate; 5: Angelol-A; 6: Angelol-B. Coumarins were incubated with Caco-2 cells for 90 min. The values presented M±SD (n=3)."

Table 1

Papp values of 6 coumarins from bidirections at different concentrations ($\bar{x}$±s)"

Compounds Papp AP→BL
(×10-5 cm/s)
Papp BL→AP
(×10-5 cm/s)
Papp AP→BL/PappBL→AP AT (%) TRR (%)
Umbelliferone
25 (μmol/L) 2.470±0.278 2.980±0.433 0.83 3.31±0.17 83.58±6.35
50 (μmol/L) 2.638±0.148 3.640±0.297 0.72 2.32±0.49 81.66±1.22
100 (μmol/L) 2.929±0.125 3.517±0.158 0.83 1.01±0.11 84.68±0.82
Average 2.679±0.263 3.381±0.410 0.79 2.21±1.06 83.31±3.52
Osthole
25 (μmol/L) 1.553±0.331 0.975±0.125 1.59 30.05±0.01 52.30±0.01
50 (μmol/L) 1.385±0.116 0.898±0.122 1.54 36.64±0.41 55.95±3.06
100 (μmol/L) 0.955±0.237 0.819±0.152 1.17 41.75±1.02 55.99±0.38
Average 1.306±0.324 0.898±0.134 1.46 36.15±5.87 54.75±2.75
Columbianadin
25 (μmol/L) 0.630±0.107 0.719±0.078 0.88 56.89±2.53 59.27±4.65
50 (μmol/L) 0.497±0.045 0.425±0.029 1.17 55.96±2.09 53.58±3.19
100 (μmol/L) 0.657±0.016 0.384±0.058 1.71 48.86±2.89 46.57±4.12
Average 0.595±0.086 0.510±0.183 1.17 53.90±4.39 53.14±6.52
Columbianetin acetate
25 (μmol/L) 0.253±0.026 0.173±0.048 1.46 6.51±0.53 8.39±0.12
50 (μmol/L) 0.343±0.021 0.252±0.075 1.36 2.39±0.29 10.25±0.12
100 (μmol/L) 0.284±0.011 0.241±0.020 1.18 5.11±2.15 8.57±0.81
Average 0.293±0.041 0.222±0.025 1.33 4.92±2.13 9.07±0.98
Angelol-A
25 (μmol/L) 1.976±0.065 1.577±0.177 1.25 5.93±0.59 86.89±0.69
50 (μmol/L) 1.639±0.088 0.968±0.030 1.69 3.74±0.36 71.06±2.61
100 (μmol/L) 0.980±0.092 1.065±0.018 0.92 2.37±0.11 74.21±0.63
Average 1.532±0.444 1.203±0.280 1.27 4.01±1.79 77.39±7.38
Angelol-B
25 (μmol/L) 1.496±0.128 0.656±0.042 2.28 3.11±0.70 74.34±7.34
50 (μmol/L) 1.604±0.036 0.837±0.042 1.92 2.28±0.59 67.10±1.70
100 (μmol/L) 1.140±0.279 0.769±0.060 1.48 1.83±0.06 50.25±8.07
Average 1.413±0.243 0.754±0.092 1.87 2.41±0.73 63.94±12.08

Figure 4

Time kinetics curve of angelol-B transported from AP side to BL side in the Caco-2 cell monolayer model The final concentration of angelol-B was 50 μmol/L. The values presented M±SD (n=3)."

Figure 5

The Papp values of angelol-B presented with EBSS, iodoacetamide or MK571 in Caco-2 cell monolayer model The values presented M±SD (n=3)."

[1] The Pharmacopoeia Commission of PRC. Pharmacopoeia of People's Republic of China(part 1)[M]. Beijing: Chemical Industry Press, 2005: 185
国家药典委员会. 中华人民共和国药典(第一部)[M]. 北京: 化学工业出版社, 2005: 185
[2] Zhang CY, Zhang BG, Yang XW . Studies on the chemical constituents of the root of Angelica pubescens f. biserrata[J]. Jie Fang Jun Yao Xue Xue Bao, 2007,23(4):241-245
doi: 10.3969/j.issn.1008-9926.2007.04.001
张才煜, 张本刚, 杨秀伟 . 独活化学成分的研究[J]. 解放军药学学报, 2007,23(4):241-245
doi: 10.3969/j.issn.1008-9926.2007.04.001
[3] Kwon YS, Kobayashi A, Kajiyama S , et al. Antimicrobial constituents of Angelica dahurica roots[J]. Phytochemistry, 1997,44(5):887-889
doi: 10.1016/S0031-9422(96)00634-6 pmid: 9115693
[4] Guh JH, Yu SM, Ko FN , et al. Antiproliferative effect in rat vascular smooth muscle cells by osthole, isolated from Angelica pubescens[J]. Eur J Pharmacol, 1996,298(2):191-197
doi: 10.1016/0014-2999(95)00812-8
[5] Yang XW, Xu B, Ran FX , et al. Inhibitory effects of 11 coumarin compounds against growth of human bladder carcinoma cell line E-J in vitro[J]. Zhong Xi Yi Jie He Xue Bao, 2007,5(1):56-60
doi: 10.3969/j.issn.1673-4890.2006.11.003
杨秀伟, 徐波, 冉福香 , 等. 11种香豆素类化合物对人膀胱癌细胞系E-J细胞株生长抑制活性的筛选[J]. 中西医结合学报, 2007,5(1):56-60
doi: 10.3969/j.issn.1673-4890.2006.11.003
[6] Yang XW, Xu B, Wu J , et al. Inhibitory effects of 40 coumarins compounds against growth of human nasopharyngeal carcinoma cell Line KB and human leukemia cell line HL-60 in vitro[J]. Zhongguo Xian Dai Zhong Yao, 2006,8(10):8-13
杨秀伟, 徐波, 吴军 , 等. 40种香豆素类化合物对人鼻咽癌细胞株KB和人白血病细胞株HL-60细胞生长抑制活性的筛选[J]. 中国现代中药, 2006,8(10):8-13
[7] Yang XW, Xu B, Ran FX , et al. Inhibitory effects of 40 coumarins compounds against growth of human gastric carcinoma cell line BGC and human hepatic carcinoma cell line BEL-7402 in vitro[J]. Zhongguo Xian Dai Zhong Yao, 2006,8(11):7-9, 24
杨秀伟, 徐波, 冉福香 , 等. 40种香豆素类化合物对人胃癌细胞株BGC和人肝癌细胞株BEL-7402细胞生长抑制活性的筛选[J]. 中国现代中药, 2006,8(11):7-9, 24
[8] Yang XW, Yang XD, Wang Y , et al. Establishment of Caco-2 cell monolayer model and standard operation procedure for assessing intestinal absorption of chemical components of traditional Chinese medicine[J]. Zhong Xi Yi Jie He Xue Bao, 2007,5(6):634-641
杨秀伟, 杨晓达, 王莹 , 等. 中药化学成分肠吸收研究中Caco-2细胞模型和标准操作规程的建立[J]. 中西医结合学报, 2007,5(6):634-641
[9] Yee S . In vitro permeability across Caco-2 cells(colonic) can predict in vivo(small intestinal) absorption in man--fact or myth[J]. Pharm Res, 1997,14(6):763-766
doi: 10.1023/A:1012102522787
[10] Gao YR, Zhang L, Zhang DS , et al. The preparation of inclusion complex of osthol and determination of bioavailability in rabbit[J]. Zhongguo Yi Yuan Yao Xue Za Zhi, 2005,25(12):1143-1146
doi: 10.3321/j.issn:1001-5213.2005.12.019
高永荣, 张力, 张丹参 , 等. 蛇床子素包合物的制备及其兔体内生物利用度测定[J]. 中国医院药学杂志, 2005,25(12):1143-1146
doi: 10.3321/j.issn:1001-5213.2005.12.019
[11] Konishi Y, Kobayashi S, Shimizu M . Transepithelial transport of p-coumaric acid and gallic acid in Caco-2 cell monolayers[J]. Biosci Biotechnol Biochem, 2003,67(11):2317-2324
doi: 10.1271/bbb.67.2317
[12] Bartels H, Korsiak E , Daniel H.Transport activity of the MDR1-gene product in monolayers of Caco-2 cells[J].Zeitschrift fur Gastroenterologie, 1994, 32: 15-18
[13] Artursson P, Palm K, Luthman K . Caco-2 monolayers in experimental and theoretical predictions of drug transport[J]. Adv Drug Deliv Rev, 2001,46(1-3):27-43
doi: 10.1016/j.addr.2012.09.005 pmid: 11259831
[1] Pragya Sharma, Sonali Batra, Ashwani Kumar, Anupam Sharma. In vivo antianxiety and antidepressant activity of Murraya paniculata leaf extracts. Journal of Integrative Medicine, 2017, 15(4): 320-325.
[2] Hai-sheng You , Hai-feng Zhang, Ya-lin Dong, Si-ying Chen, Mao-yi Wang, Wei-hua Dong, Jian-Feng Xing. Absorption and transportation characteristics of scutellarin and scutellarein across Caco-2 monolayer model. Journal of Chinese Integrative Medicine, 2010, 8(9): 863-869.
[3] Yan Zheng, Xiu-wei Yang. Absorption and transport of pachymic acid in the human intestinal cell line Caco-2 monolayers. Journal of Chinese Integrative Medicine, 2008, 6(7): 704-710.
[4] Xiu-wei Yang, Xiao-da Yang, Ying Wang, Lian Ma, Yue Zhang, Xiao-gai Yang, Kui Wang. Establishment of Caco-2 cell monolayer model and standard operation procedure for assessing intestinal absorption of chemical components of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2007, 5(6): 634-641.
[5] Xiu-wei Yang, Bo Xu, Fu-xiang Ran, Rui-qing Wang, Jun Wu, Jing-rong Cui. Inhibitory effects of 11 coumarin compounds against growth of human bladder carcinoma cell line E-J in vitro. Journal of Chinese Integrative Medicine, 2007, 5(1): 56-60.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] Chan Chen, Yong-mei Meng , Peng Zhang, Juan Wang, Hui-hui Zhao, Shu-zhen Guo , Wei Wang. Diagnosis and treatment rule of traditional Chinese medicine for syndrome factors of chronic congestive heart failure: a study based on Shannon entropy method. Journal of Chinese Integrative Medicine, 2010, 8(11): 1080-1084
[2] Liang-ping Hu, Xiao-lei Bao. Three-factor designs unable to examine the interactions (Part 2). Journal of Chinese Integrative Medicine, 2012, 10(11): 1229-1232
[3] Ming-qing Wei , Jin-zhou Tian , Jing Shi , Fu-yun Ma , Ying-chun Miao , Yong-yan Wang. Effects of Chinese medicine for promoting blood circulation and removing blood stasis in treating patients with mild to moderate vascular dementia: a randomized, double-blind and parallel-controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(11): 1240-1246
[4] Ji Chen, Qin Pan, Li-shuang Ye, Jia-ling Huang. English translation of cultural aspects of the titles of traditional Chinese medicine classics based on the skopos theory. Journal of Chinese Integrative Medicine, 2012, 10(11): 1316-1320
[5] Oluwole Busayo Akinola. Should complementary and alternative medicine familiarisation modules be taught in African medical schools?. Journal of Chinese Integrative Medicine, 2011, 9(11): 1165-1169
[6] Xiang-yu Kong, Yuan Hao, Tai-xiang Wu, Yan-ming Xie. Adverse drug reactions or adverse events of Chaihu Injection: A systematic review. Journal of Chinese Integrative Medicine, 2010, 8(12): 1124-1132
[7] Yong-mei Liu, Rui-hua Liu, Wen-jun Liu, Li Liu , Zhi-kui Wu, Yu-ying Chen. Effects of Chinese herbal medicine Yiqi Huoxue Formula on TGF-p/smad signal transduction pathway and connective tissue growth factor in rats with renal interstitial fibrosis. Journal of Chinese Integrative Medicine, 2010, 8(12): 1165-1173
[8] Mei-na Ye, Hong-feng Chen, Rui-juan Zhou, Ming-juan Liao. Effects of Astragalus polysaccharide on proliferation and Akt phosphorylation of the basal-like breast cancer cell line. Journal of Chinese Integrative Medicine, 2011, 9(12): 1339-1346
[9] Jie Wang, Yan-wei Xing, Jian-xin Chen, Qing-yong He, Yong-hong Gao, Zun Li. Characteristics of coronary arteriography and traditional Chinese medicine syndrome of 1 069 patients with coronary artery disease. Journal of Chinese Integrative Medicine, 2008, 6(2): 148-152
[10] Ru-jiang Li, Shu-dong Qiu, Hong-xia Chen, Li-rong Wang. Immunomodulatory effects of Astragalus polysaccharide in diabetic mice. Journal of Chinese Integrative Medicine, 2008, 6(2): 166-170