Intestinal Transport of 3,6′-Disinapoylsucrose, A Major Active Component of Polygala tenuifolia, Using Caco-2 Cell Monolayer and In Situ Rat Intestinal Perfusion Models

Abstract

3,6'-Disinapoylsucrose is a major active component of the herb Polygala tenuifolia which has long been used for relieving tranquilization, uneasiness of the mind, and improving learning and memory. Our previous study found that 3,6'-disinapoylsucrose had a very low oral bioavailability. Its mechanisms of absorption in the small intestine have so far been unclear. In the present study, the absorption mechanisms of 3,6'-disinapoylsucrose were investigated by using the Caco-2 cell monolayer and in situ rat intestinal perfusion models. The 3,6'-disinapoylsucrose concentration was determined by an LC/MS/MS method. In a Caco-2 cell transport study, the results showed that 3,6'-disinapoylsucrose had very limited intestinal permeability with average apparent permeability coefficient values around (1.11-1.34) × 10(-7) cm/s from the apical (A) to the basolateral (B) side and (1.37-1.42) × 10(-7) cm/s from B to A, at concentrations of 5, 20, and 33 µM. No concentration dependence in the 3,6'-disinapoylsucrose transport was observed. The apparent permeability coefficient value of 3,6'-disinapoylsucrose (5 µM) from A to B greatly increased to 4.49 × 10(-7) and 1.81 × 10(-7) cm/s, respectively, when the cells were preincubated with EDTA (17 mM) and sodium caprate (5.14 mM). No significant effect on the 3,6'-disinapoylsucrose transport by the inhibitors including verapamil, cyclosporine A, and sodium azide was observed. Similar results were found in the small intestinal perfusion study. The apparent permeability coefficient value of 3,6'-disinapoylsucrose greatly increased from 3.97 × 10(-6) to 23.4 × 10(-6) and 20.0 × 10(-6) cm/s in the presence of EDTA (17 mM) and sodium caprate (5.14 mM), respectively, in perfusion buffer. An in vitro stability evaluation of 3,6'-disinapoylsucrose in the gastrointestinal tract showed that it was relatively stable both in the stomach and small intestine contents, while it was found to be more instable in the colon contents. All of the above results indicate that 3,6'-disinapoylsucrose might be transported across the intestinal mucosa by paracellular passive penetration and paracellular enhancers could increase the intestinal permeability of this compound and thus slightly improve its oral bioavailability.

Transport of Corilagin, Gallic Acid, and Ellagic Acid from Fructus Phyllanthi Tannin Fraction in Caco‐2 Cell Monolayers

“…Both SDS and PLL greatly improved the transport of corilagin, GA, and EA. Other tight junction modulators such as EDTA and sodium caprate could enhance the transport of 3,6′-disinapoylsucrose [ 28 ]. These results further suggested that the paracellular transport appeared to be main limiting factor for the uptake of three compounds in Caco-2 cell monolayers.…”

“…Then, the DMEM culture medium was replaced with 150 μ L HBSS and incubated with different concentrations of fractions for 4 h. The concentration of PTF was ranged from 0.25 to 2 mg/mL. Thereafter, the HBSS was removed and the monolayers were incubated with 200 μ L of DMEM culture medium containing MTT 500 μ g/L for 4 h. Finally, the medium was removed, the formazan salt crystals that remained were dissolved with 150 μ L of DMSO and shaken for 10 min, and the absorbance was measured at 570 nm using microplate reader (Molecular Devices, Sunnyvale, CA, USA) [ 28 ]. The cell inhibitory rate was calculated as follows: inhibitory rate (%) = (A570 control − A570 sample )/(A570 control − A570 blank ) × 100%.…”

“…It is essential to find out why the low absorption of oral PTF occurred and to identify suitable pharmaceutical methods to improve the absorption of PTF and further enhance the anticancer activities. P-gp inhibitor (verapamil) [ 21 , 22 ] and MRPs inhibitor (indomethacin) [ 24 , 25 ] were employed to investigate whether the transport was affected by the efflux transporters, OATP inhibitor (indomethacin) [ 26 , 27 ] and SGLT1 inhibitor (phloridzin) [ 21 , 28 ] were chosen to study the effect of absorption transporters, and tight junction modulators (sodium dodecyl sulfate (SDS) and poly-l-lysine (PLL)) [ 29 , 30 ] were used to evaluate the role of paracellular permeability in the transport of these three compounds from PTF.…”

Exploring the compatibility mechanism of ShengDiHuang Decoction based on the in situ single‐pass intestinal perfusion model

“…To date, the in vitro Caco‐2 cell model, single‐pass intestinal perfusion (SPIP) in situ model, and the everted intestinal sac in vivo model have been used frequently to study intestinal transport mechanisms (Chen et al, ; Hewei Li et al, ; Zhai et al, ). The SPIP in situ model provides optimal oral conditions in vivo for investigating the absorption potential of drugs in the intestine, and is used more commonly than other models (Bohets et al, ; Sutton, Rinaldi, & Vukovinsky, ).…”

“…Malabsorption of compounds, due to low transport function, can effectively be improved by using a tight junction regulator (Bohets et al, ; Sutton et al, ). In many recent studies, verapamil was used as an inhibitor of P‐gp, indomethacin as an inhibitor of MRP2, reserpine as an inhibitor of BCRP, and EDTA as a tight junction regulator (Chen et al, ; Duan et al, ; Li et al, ). In our previous work, the mechanism for the synergistic effect of SDHD was clarified at the component–effect level.…”