Intestinal transport of sophocarpine across the Caco‐2 cell monolayer model and quantification by LC/MS

Abstract

Sophocarpine is a biologically active component obtained from the foxtail-like sophora herb and seed that is often orally administered for the treatment of cancer and chronic bronchial asthma. The aim of this study was to develop a rapid and specific LC/MS method for the determination of sophocarpine and to explore its transcellular transport mechanism across the Caco-2 (the human colon adenocarcia cell lines) monolayer cell transwell model. Caco-2 cells were seeded on permeable polycarbonate membranes and incubated for 21 days. Before the experiment, the trans-epithelial electric resistance, integrity and alkaline phosphatase activity of the Caco-2 monolayers were verified and used in subsequent experiments. In the Caco-2 model constructed, many influencing factors were investigated, including time, concentration, pH and different protein inhibitors. The results suggested that sophocarpine was transported mainly by passive diffusion. The flux of sophocarpine was time- and concentration-dependent, and the pH also had an effect on its transportation. The PappBA was higher than PappAB , indicating that a polarized transport might exist for sophocarpine. MK-571 and reserpine, inhibitors of the multidrug resistance associated protein 2 and the breast cancer resistance protein, decreased the efflux of sophocarpine, while verapamil had no effect on its transport. These results revealed that sophocarpine is absorbed mainly by passive diffusion, and that a carrier-mediated mechanism is also involved in the transport of sophocarpine.

Absorption and Metabolism Characteristics of Triptolide as Determined by a Sensitive and Reliable LC-MS/MS Method

“…Caco-2 cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). Caco-2 cell transport experiment was performed according to the previously reported method [18,19]. In brief, Caco-2 cells were cultured in DMEM high glucose media containing 15% FBS, 1% NEAA and 100 U·mL −1 penicillin and streptomycin.…”

Biological activities and pharmacokinetics of aconitine, benzoylaconine, and aconine after oral administration in rats

“…[26] In brief, Caco-2 cells were cultured in MEM medium containing 15% FBS, 1% NEAA and 100 U·mL -1 penicillin and streptomycin, and maintained at 37°C in a humidified, 5% CO 2 incubator. For transportation study, cells at passage 30-40 were seeded on transwell polycarbonate insert filters (1.12 cm 2 surface, 0.4 μm pore size, 12 mm diameter) in 12-well plates at a density of 1 × 10 5 cells·cm -2…”

Enhanced Bioavailability by Orally Administered Sirolimus Nanocrystals

“…That is, the cell layer takes the drug from the GIT and transports it to the blood/mesenteric lymph . In contrast, when drug HBSS buffer was added to the BL side and blank HBSS buffer was added to the AP side, the process of reversing the blood side of the animal into the intestinal lumen was simulated. , The efflux rate of SRL NCs, calculated as P app, BL‑AP / P app, AP‑BL , was significantly different from that of SRL (Figure C). The reason may be that the drug became smaller and the surface area increased after nanocrystallization, so transport across the monolayer was significantly enhanced .…”