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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

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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
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