• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1996.04a
• /
• pp.119-130
• /
• 1996

We review the factors which regulate the depth and composition of the human airway surface liquid (ASL). These include secretion from airway submucosal glands, ion and fluid transport across the surface epithelium, goblet cell discharge, surface tension and transepithelial gradients in osmotic and hydrostatic pressure. We describe recent experiments in which we have used low temperature scanning electron microscopy of rapidly frozen specimens to detect changes in depth of ASL in response to submucosal gland stimulation. We also present preliminary data in which X-ray microanalysis of frozen specimens has been used to determine the elemental composition of ASL.

• Journal of Pharmaceutical Investigation
• /
• v.34 no.6
• /
• pp.483-489
• /
• 2004

Fexofenadine HCl is non-sedating histamine H1 receptor antagonist that can be used for the treatment of seasonal allergic rhinitis. The objective of this study was to investigate whether the carriers of deformable liposomes can enhance the transepithelial permeability of fexofenadine HCl across the in vitro ALI human nasal monolayer model. Characterization of this model was achieved by bioelectric measurements and morphological studies. The passage 2 and 3 of cell monolayers exhibited the TEER value of $2852\;{\pm}\;482\;ohm\;{\times}\;cm^2$ on 11 days of seeding and maintained high TEER value for 5 days. The deformable liposome of fexofenadine HCl was prepared with phosphatidylcholine (PC) and cholic acid using extruder method. The mean particle size was about 200 nm and the maximum entrapment efficiency of 33.0% was obtained in the formulation of 1% PC and $100\;{\mu}g/ml$ fexofenadine HCl. The toxicity of the deformable liposome to human nasal monolayers was evaluated by MTT assay and TEER value change. MTT assay showed that it has no toxic effect on the nasal epithelial cells in 2-hour incubation when the PC concentration was below 1%. However, deformable liposome could not enhance the transepithelial permeability $(P_{app})$ and cellular uptake of fexofenadine HCl. In conclusion, the in vitro model could be used in nasal drug transport studies and evaluation of transepithelial permeability of formulations.

• The Korean Journal of Physiology
• /
• v.30 no.1
• /
• pp.53-62
• /
• 1996

Transepithelial transport of tetraethylammonium (TEA) was studied in monolayers of opossum kidney cells cultured on permeable membrane filters. $[^{14}C]-TEA$ was transported across the OK cell monolayer from basolateral to apical side by a saturable process which can be stimulated by acidification of the apical medium. The apparent Michaelis-Menten constant $(K_{m})$ and the maximum velocity$(V_{max})$ for the transport were $41\;{\mu}M$ and 147 pmole/ mg protein/ min, respectively. The transport was significantly inhibited by unlabelled TEA, amiloride, cimetidine, choline, and mepiperphenidol added to the basolateral side at 1 mM and was slightly inhibited by 5 mM $N_{1}-methylnicotinamide\;(NMN).$ Unlabelled TEA added to the apical side stimulated the $basolateral-to-apical\;{^{14}C}-TEA$ transport, suggesting that the TEA self-exchange mechanism was involved at the apical membrane. Sulfhydryl reagents such as ${\rho}-chloromercuribenzoic\;acid\;(PCMB)\;and \;{\rho}-chloro-mercuribenzene\;sulfonate \;(PCMBS)$ and carboxyl reagents such as N,N'-dicyclohexylcarbodiimidem (DCCD) and N-ethoxy-carbonyl-2-ethoxy-1,2-dihydro-quinoline(EEDQ) inhibited the TEA transport at both the basolateral and apical membranes of the OK cell monolayer. These results suggest that OK cell monolayers possess a vectorial transport system for organic cations which is similar to that for organic cation secretion in the renal proximal tubule.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.234.3-235
• /
• 2003

The aim of this study is to evaluate the permeability of PEG-conjugated salmon calcitonin (sCT) across monolayers of Caco-2 cells that represent a model of the intestinal barrier. Caco-2 cells were grown to confluency on a permeable polycarbonate membrane to permit transport through it. Permeability experiments were performed with native-sCT and PEG-conjugated sCT (PEG M.W. 2000) at various concentrations (5uM, 10uM, 25uM, 50uM, 100uM) in the apical to basolateral direction. The barrier properties were assessed by detecting transport of markder molecules ($^3$H-mannitol) and by measuring transepithelial electrical resistance (TEER). (omitted)

• The Korean Journal of Physiology
• /
• v.10 no.1
• /
• pp.35-40
• /
• 1976

In an attempt to examine the effect of ginseng on sodium transport across the biological membrane, we have studied effects of ginseng alcohol extract on the short-circuit current(SCC) and the $Na^+-K^+$-activated ATPase activity in isolated frog skin preparations. 1. Ginseng alcohol extract applied to the mucosal surface of the frog skin significantly increased SCC at low concentration($1{\sim}10mg%$) but decreased SCC at higher concentration($50{\sim}250mg%$). 2. Similarly, when the drug was added to the serosal bathing medium, the SCC was stimulated at low doses($5{\sim}25mg%$) and inhibibited at high doses($50{\sim}250mg%$). 3. $Na^+-K^+$-activated ATPase activity of the frog skin epidermal homogenate was significantly inhibited by ginseng alcohol extract, the effect being proportional to the concentration of the drug in the incubation mixture. These results may suggest that a low dose of ginseng alcohol extrat enhances the transepithelial sodium transport probably by increasing the permeability of outer membrane of the transporting cell to sodium ion, whereas a high dose of drug reduces the sodium transport primarly by inhibiting $Na^+-K^+$ ATPase mediated active transport step.

• Archives of Pharmacal Research
• /
• v.25 no.1
• /
• pp.86-92
• /
• 2002

The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested . The cytotoxicity of these enhancers was also examined. The enhancing effects by Quillaja saponin, diponin glycyrrhizinate, $18{\beta}-glycyrrhetinic$ acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, $18{\beta}-glycyrrhetinic$ acid arid taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds . It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium.

• Applied Microscopy
• /
• v.23 no.1
• /
• pp.1-14
• /
• 1993

The present study was carried out to analyze the evidence of membrane recycling, and the regulation of cellular transport by dynamic changes in apical membrane area that functionally interacts with the number of cytoplasmic vesicles. Under scanning electron micrographs, turtle bladder mucosa contain three main type of cells; granular cells and carbonic anhydrase (CA)-rich cells, deviding into a and b type of epithelial cell. The granular cell is the majority cell type of the mucosa comprising 80% of the total cell number. The remaining 20% of the cells are characteristically rich in carbonic anhydrase. Uptake of HRP was detected in the most vacuoles or tubulovesicles in both type of CA-rich cells in the turtle bladder, indicating that the part of plasma membrane was internalized in the apical cytoplasmic vacuoles. It seems quite likely that CA-rich cells possess intracellular vesicles carrying proton pumps which are recycling back to the apical plasma membrane. In turtle bladder, the granular cells actively secrete large quantities of mucin and other proteins by an exocytotic mechanism in an apparently constitutive fashion. The possibility that bladder epithelial cells secrete mucin via a regulated secretory pathway has not been rigorously examined and much is still to be determined about these issues from this cell type.

• Journal of Pharmaceutical Investigation
• /
• v.32 no.1
• /
• pp.21-26
• /
• 2002

The primary culture of human nasal epithelial cell monolayer was performed on a Transwell. The effect of various factors on the tight junction formation was observed in order to develop an in vitro experimental system for nasal transport studies. Human nasal epithelial cells, collected from human normal inferior turbinates, were plated onto diverse inserts. After 4 days, media of the apical surface was removed for air-liquid interface (ALI) culture. Morphological characteristics was observed by transmission electron microscopy (TEM). A polyester membrane of $0.4\;{\mu}m$ pore size was determined as the most effective insert based on the change in the transepithelial electric resistance (TEER) value as well as the $^{14}C-mannitol$ transport study. The ALI method was effective in developing the tight junction as observed in the further increase in the TEER value and reduction in the permeability coefficient $(P_{app})$ of $^{14}C-mannitol$ transport. Results of the transport study of a model drug, budesonide, showed that the primary culture system developed in this study could be further developed and applied for in vitro nasal transport studies.

• Journal of Ginseng Research
• /
• v.45 no.1
• /
• pp.66-74
• /
• 2021

Background: Abnormal chloride (Cl-) transport has a detrimental impact on mucociliary clearance in both cystic fibrosis (CF) and non-CF chronic rhinosinusitis. Ginseng is a medicinal plant noted to have anti-inflammatory and antimicrobial properties. The present study aims to assess the capability of red ginseng aqueous extract (RGAE) to promote transepithelial Cl- secretion in nasal epithelium. Methods: Primary murine nasal septal epithelial (MNSE) [wild-type (WT) and transgenic CFTR-/-], fisher-rat-thyroid (FRT) cells expressing human WT CFTR, and TMEM16A-expressing human embryonic kidney cultures were utilized for the present experiments. Ciliary beat frequency (CBF) and airway surface liquid (ASL) depth measurements were performed using micro-optical coherence tomography (μOCT). Mechanisms underlying transepithelial Cl- transport were determined using pharmacologic manipulation in Ussing chambers and whole-cell patch clamp analysis. Results: RGAE (at 30㎍/mL of ginsenosides) significantly increased Cl- transport [measured as change in short-circuit current (ΔISC = ㎂/㎠)] when compared with control in WT and CFTR-/- MNSE (WT vs control = 49.8±2.6 vs 0.1+/-0.2, CFTR-/- = 33.5±1.5 vs 0.2±0.3, p < 0.0001). In FRT cells, the CFTR-mediated ΔISC attributed to RGAE was small (6.8 ± 2.5 vs control, 0.03 ± 0.01, p < 0.05). In patch clamp, TMEM16A-mediated currents were markedly improved with co-administration of RGAE and uridine 5-triphosphate (8406.3 +/- 807.7 pA) over uridine 5-triphosphate (3524.1 +/- 292.4 pA) or RGAE alone (465.2 +/- 90.7 pA) (p < 0.0001). ASL and CBF were significantly greater with RGAE (6.2+/-0.3 ㎛ vs control, 3.9+/-0.09 ㎛; 10.4+/-0.3 Hz vs control, 7.3 ± 0.2 Hz; p < 0.0001) in MNSE. Conclusion: RGAE augments ASL depth and CBF by stimulating Cl- secretion through CaCC, which suggests therapeutic potential in both CF and non-CF chronic rhinosinusitis.

• Journal of Life Science
• /
• v.20 no.12
• /
• pp.1784-1791
• /
• 2010

In this study, we investigated the effects of ethyl alcohol extracts of Hizikia fusiforme (EHF) on the correlation between tightening of tight junctions (TJs) and anti-invasive activity in human gastric adenocarcinoma AGS cells. Inhibitory effects of EHF on cell proliferation, motility, and invasiveness were found to be associated with increased tightness of the TJs, which was demonstrated by an increase in transepithelial electrical resistance. Activities of matrix metalloprotease (MMP)-2 and -9 in AGS cells were dose-dependently inhibited by treatment with EHF, and this was also correlated with a decrease in expression of their mRNA and proteins; however, tissue inhibitor of metalloproteinase (TIMP)-1 and -2 mRNA levels were increased. Additionally, immunoblotting results indicated that EHF repressed the levels of claudin proteins (claudin-1, -3, and -4), major components of TJs that play key roles in control and selectivity of paracellular transport. Furthermore, EHF decreased expression of insulin such as growth factor-1 receptor proteins, while concurrently increasing that of thrombospondin-1 and E-cadherin. In conclusion, these results suggest that EHF treatment may inhibit tumor cell motility and invasion, and therefore act as a dietary source to decrease the risk of cancer metastasis.