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Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (7): 704-710.doi: 10.3736/jcim200800709

• Original Experimental Research • Previous Articles     Next Articles

Absorption and transport of pachymic acid in the human intestinal cell line Caco-2 monolayers

Yan Zheng, Xiu-wei Yang()   

  1. State Key Laboratory of Natural and Biomimetic Drugs, Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China
  • Received:2007-11-14 Online:2008-07-20 Published:2008-07-15
  • Contact: YANG Xiu-wei E-mail:xwyang@bjmu.edu.cn

Objective: To study the absorption and transport of pachymic acid (PA) isolated from the sclerotium of Poria cocos (Schw.) Wolf. in human intestinal epithelium.
Methods: By using Caco-2 (the human colonic adenocarcinoma cell lines) cell monolayers as an intestinal epithelial cell model, the permeability of PA was studied from apical side (AP side) to basolateral side (BL side) or from BL side to AP side. The PA was measured by reversed-phase high performance liquid chromatography coupled with UV detector at maximum absorption wavelength of 210 nm. Transport parameters and apparent permeability coefficients (Papp) were then calculated and compared with those of propranolol and atenolol, which were the transcellular transport markers for high and poor permeability respectively.
Results: The Papp values of PA were (9.50±2.20) 10 –7 cm/s from AP side to BL side, and (11.30±5.90) 10 –7 cm/s from BL side to AP side, respectively. Under the condition of this experiment, the Papp values were 1.45×10 –5 cm/s for propranolol and 4.22×10 –7 cm/s for atenolol.
Conclusion: PA is transported through the Caco-2 cell monolayer in a concentration-dependent manner and the transport was linear with time. The absorption in apical to basolateral direction and secretion in basolateral to apical direction were poor and their Papp values were comparable to atenolol. Besides passive diffusion of PA, ATP is partially involved in its transport.

Key words: sclerotium of Poria cocos (Schw.) Wolf., pachymic acid, Caco-2 cell monolayer, intestinal absorption

CLC Number: 

  • R285.5

Figure 1

Chemical structure of PA"

Figure 2

The apical-to-basolateral (■) and basolateral to apical (○) transport rates of the PA at different concentrations The incubation time was up to 180 minutes and all experiments were carried out in triplicate ($\bar{x}$±s )."

"

Compound Direction Papp ($\bar{x}$±s, ×10–6 cm/s) Papp AP→BL/Papp BL→AP
20 μmol/L 40 μmol/L 50 μmol/L Average
PA AP→BL 1.20±0.62 0.89±0.11 0.77±0.50 0.95±0.22 0.84
BL→AP 1.01±0.04 1.76±0.22 0.61±0.17 1.13±0.59

Figure 3

Kinetics curve of PA transport in Caco-2 cell monolayers from apical to basolateral direction at 45 μmol/L"

Figure 4

Influence of sodium azide (SA) at different concentrations on the Papp values of PA from both directions by Caco-2 cell monolayers The incubation time was 180 minutes and all experiments were carried out in triplicate. Data are the x±s. *P<0.05, vs untreated group."

Figure 5

Influence of verapamil (VP) at different concentrations on the Papp values of PA from both directions by Caco-2 cell monolayers The incubation time was 180 minutes and all experiments were carried out in triplicate. Data are the $\bar{x}$±s. *P<0.05, **P<0.01, vs untreated group."

Figure 6

Influence of probenicid (PR) at different concentrations on the Papp values of PA from both directions by Caco-2 cell monolayers The incubation time was 180 minutes and all experiments were carried out in triplicate. Data are the $\bar{x}$±s. *P< 0.05, vs untreated group."

"

C0 of PA (μmol/L) Detected part Concentration and total recovery
AP→BL BL→AP
20 A 6.97±9.20 0.24±0.01
B 0.29±0.15 5.32±1.82
C 0.01±0.00 -
R 72.80±1.49 18.56±6.10
40 A 14.51±9.54 0.85±1.07
B 0.43±0.05 18.90±3.06
C 0.10±0.00 0.35±0.00
R 75.33±47.99 33.53±6.89
50 A 9.18±1.00 0.25±0.07
B 0.31±0.20 34.13±3.19
C 0.69±0.18 0.34±0.13
R 40.69±5.52 46.24±4.52
[1] Chinese Pharmacopoeia Commission. Pharmacopoeia of People's Republic of China(Vol. I)[M]. Beijing: Chemical Industry Press,2005: 166
国家药典委员会. 中华人民共和国药典(一部)[M]. 北京: 化学工业出版社,2005: 166
[2] Zheng Y, Yang XW. Modern study on Poria cocos. In: Huang TK. Frontier for drug research (Vol. 2004). Beijing: China Medico-Pharmaceutical Science & Technology Publishing House. 2005: 300-321. Chinese
郑艳, 杨秀伟 . 茯苓的现代研究概况. 见: 黄泰康. 药学前沿 (2004年卷). 北京: 中国医药科技出版社. 2005: 300-321
[3] Nukaya H, Yamashiro H, Fukazawa H , et al. Isolation of inhibitors of TPA-induced mouse ear edema from Hoelen, Poria cocos[J]. Chem Pharm Bull (Tokyo), 1996,44(4):847-849
doi: 10.1248/cpb.44.847
[4] Tai T, Akita Y, Kinoshita K , et al. Anti-emetic principles of Poria cocos[J]. Planta Med, 1995,61(6):527-530
doi: 10.1055/s-2006-959363 pmid: 8824947
[5] Ukiya M, Akihisa T, Tokuda H , et al. Inhibition of tumor-promoting effects by poricoic acids G and H and other lanostane-type triterpenes and cytotoxic activity of poricoic acids A and G from Poria cocos[J]. J Nat Prod, 2004,65(4):462-465
[6] Gan LSL , Thakker DR.Application of the Caco-2 model in the design and development of orally active drugs: Elucidation of biochemical and physical barriers posed by the intestinal epithelium[J].Adv Drug Deliv Rev, 1997, 23: 77-98
doi: 10.1016/S0169-409X(96)00427-9
[7] Yang XW, Guo QM, Wang Y , et al. Intestinal permeability of antivirus constituents from the fruits of Eucalyptus globulus Labill. in Caco-2 cell model[J]. Bioorg Med Chem Lett, 2007,17(4):1107-1111
doi: 10.1016/j.bmcl.2006.11.021
[8] Tian XJ, Yang XD, Wang K , et al.The efflux of flavonoids morin, isorhamnetin-3-O-rutinoside and diosmetin-7-O-β-D-xylopyranosyl-(1-6)-β-D-glucopyranoside in the human intestinal cell line Caco-2[J].Pharm Res, 2006, 23: 1721-1728
doi: 10.1007/s11095-006-9030-5
[9] Tai T, Shingu T, Kikuchi T , et al.Isolation of lanostane-type triterpene acids having an acetoxyl group from sclerotia of Poria cocos[J].Phytochemistry, 1995, 40: 225-231
doi: 10.1016/0031-9422(95)00182-7
[10] Rashid Z, Basson MD . Topoisomerase II inhibition differentially modulates Caco-2 intestinal epithelial cell phenotype[J]. Biochem Biophys Res Commun, 1996,219(1):82-88
doi: 10.1006/bbrc.1996.0185
[11] 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):633-641
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):633-641
[12] Chong S, Dando SA, Morrison RA . Evaluation of biocoat intestinal epithelium differentiation environment(3-day cultured Caco-2 cells) as an absorption screening model with improved productivity[J]. Pharm Res, 1999,14(12):1835-1837
[13] Walle UK, French KL, Walgren RA , et al. Transport of genistein-7-glucoside by human intestinal CACO-2 cells: potential role for MRP2[J]. Res Commun Mol Pathol Pharmacol, 1999,103(1):45-56
[14] Yee SY.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: 763-766
[15] Zhou S, Li Y, Kestell P , et al. Determination of thalidomide in transport buffer for Caco-2 cell monolayers by high-performance liquid chromatography with ultraviolet detection[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2003,785(1):165-173
doi: 10.1016/S1570-0232(02)00911-X
[16] van der Sandt IC, Blom-Roosemalen MC, de Boer AG , et al. Specificity of doxorubicin versus rhodamine-123 in assessing P-glycoprotein functionality in the LLC-PK1, LLC-PK1:MDR1 and Caco-2 cell lines[J]. Eur J Pharm Sci, 2000,11(3):207-214
doi: 10.1016/S0928-0987(00)00097-X
[17] Versantvoort CH, Bagrij T, Wright KA , et al. On the relationship between the probenecid-sensitive transport of daunorubicin or calcein and the glutathione status of cells overexpressing the multidrug resistance-associated protein(MRP)[J]. Int J Cancer, 1995,63(6):855-862
doi: 10.1002/(ISSN)1097-0215
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