scholarly journals Intestinal Transportations of Main Chemical Compositions of Polygoni Multiflori Radix in Caco-2 Cell Model

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Jie Yu ◽  
Na Li ◽  
Pei Lin ◽  
Yunfei Li ◽  
Xiaojian Mao ◽  
...  
Keyword(s):  
Gibbs Energy ◽  
Cell Model ◽  
Cell Monolayer ◽  
Intestinal Transport ◽  
Transepithelial Transport ◽  
Chemical Compositions ◽  
Polygonum Multiflorum ◽  
Close Attention ◽  
Apparent Permeability ◽  
Cell Monolayers

Context. Polygoni Multiflori Radix (PMR) is originated from the root ofPolygonum multiflorumThunb. and used in oriental countries for centuries. However, little researches pay close attention to the absorption of its major constituents.Objective. Transepithelial transport of TSG, RL, PL, and four anthraquinones is carried out.Materials and Methods. Caco-2 cell monolayer, which represented a well-established model for the study of intestinal transport of nutrients and xenobiotics, was used in this paper.Results. The apparent permeability coefficients (Papp) in the Caco-2 cell monolayers were TSG (2.372 × 10−9) < EG (2.391 × 10−9) < EN (2.483 × 10−9) < PL (4.917 × 10−9) < RN (1.707 × 10−8) < RL (1.778 × 10−8) < AE (1.952 × 10−8). Thus, RN, RL, and AE were considered partly absorbed, while other constituents were hardly absorbed.Discussion and Conclusion. Glycosides showed poor permeabilities than aglycones. In the meantime, TSG and EN gave out poor recovery rates in this assay, which indicated that TSG and EN may accumulate or metabolise in the Caco-2 cells.In silicoprediction indicated that Gibbs energy (r=0.751,p<0.05) and heat of form (r=0.701,p<0.05) were strongly positively correlated withPapp.

scholarly journals A Miniaturized Pump Out Method for Characterizing Molecule Interaction with ABC Transporters

2019 ◽  
Vol 20 (22) ◽  
pp. 5529 ◽  
Author(s):  
Emmanuel Sevin ◽  
Lucie Dehouck ◽  
Romain Versele ◽  
Maxime Culot ◽  
Fabien Gosselet
Keyword(s):  
Abc Transporters ◽  
Detection Method ◽  
Cell Model ◽  
Early Stage ◽  
Cell Monolayer ◽  
Efflux Pumps ◽  
Apparent Permeability ◽  
Traditional Use ◽  
Molecule Design ◽  
Molecule Interaction

Characterizing interaction of newly synthetized molecules with efflux pumps remains essential to improve their efficacy and safety. Caco-2 cell line cultivated on inserts is widely used for measuring apparent permeability of drugs across biological barriers, and for estimating their interaction with efflux pumps such as P-gp, BCRP and MRPs. However, this method remains time consuming and expensive. In addition, detection method is required for measuring molecule passage across cell monolayer and false results can be generated if drugs concentrations used are too high as demonstrated with quinidine. For this reason, we developed a new protocol based on the use of Caco-2 cell directly seeded on 96- or 384-well plates and the use of fluorescent substrates for efflux pumps. We clearly observed that the new method reduces costs for molecule screening and leads to higher throughput compared to traditional use of Caco-2 cell model. This accelerated model could provide quick feedback regarding the molecule design during the early stage of drug discovery and therefore reduce the number of compounds to be further evaluated using the traditional Caco-2 insert method.

scholarly journals Transport Mechanisms of Polymannuronic Acid and Polyguluronic Acid Across Caco-2 Cell Monolayers

Pharmaceutics ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 167
Author(s):  
Yu Wang ◽  
Xu Bai ◽  
Bo Hu ◽  
Maochen Xing ◽  
Qi Cao ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Biological Activities ◽  
Intestinal Transport ◽  
Fluorescein Isothiocyanate ◽  
Endocytic Pathway ◽  
Detailed Knowledge ◽  
Intestinal Epithelial ◽  
Transport Mechanisms ◽  
Apparent Permeability ◽  
Cell Monolayers

Detailed knowledge of the intestinal transport of polymannuronic acid (PM) and polyguluronic acid (PG) is critical for understanding their biological activities. To investigate the transport in the gastrointestinal tract, PM and PG were chemically modified with tyramine and conjugated with fluorescein isothiocyanate (FITC) to synthesize FITC-PM (F-PM) and FITC-PG (F-PG) successfully. The transport mechanisms of F-PM and F-PG across the intestinal epithelial cell monolayers (Caco-2 cell monolayers) were then investigated. The results demonstrated that the transport of F-PM and F-PG into epithelial cells was time- and energy-dependent, which was mediated by the macropinocytosis pathway and the clathrin- and caveolae (or lipid raft)-mediated endocytic pathway. The transport process of F-PM and F-PG in Caco-2 cells depended on the acidification of endosomes and involved lysosomes. Tubulin mediated the transport of F-PM, but not of F-PG. Moreover, the absorption enhancer chitosan (CS) promoted the transport of F-PM and F-PG, increasing the apparent permeability coefficient (Papp) by 1.9-fold and 2.6-fold, respectively, by reversibly opening the tight junction (TJ). In summary, this study provided a comprehensive understanding of the transport of PM and PG in the small intestinal epithelial cells, which will provide a theoretical basis for the development of PM and PG with good intestinal absorption.

scholarly journals Absorption and Transport Characteristics and Mechanisms of Carnosic Acid

Biology ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1278
Author(s):  
Xuexiang Chen ◽  
Meigui Huang ◽  
Dongmei Liu ◽  
Yongze Li ◽  
Qiu Luo ◽  
...  
Keyword(s):  
Cell Model ◽  
Cell Monolayer ◽  
Scientific Basis ◽  
Carnosic Acid ◽  
Intestinal Epithelial ◽  
Apparent Permeability ◽  
Beneficial Effects ◽  
Single Cell Model ◽  
Relevant Dose ◽  
Apparent Permeability Coefficient

Carnosic acid (CA) is a phenolic diterpenoid mainly found in rosemary and sage. CA has been reported to possess health-beneficial effects in various experimental settings. Herein, a mouse experiment and Caco-2 single-cell model were used to understand the absorption and transport characteristics of CA. First, we determined the tissue distribution of CA in mice, following an oral gavage at a physiologically relevant dose. We found that CA was bioavailable systemically and present locally in the digestive tract, especially in the cecum and colon. Next, we thought to characterize the absorption and transport of CA in the Caco-2 cell monolayer model of the intestinal epithelial barrier. In the Caco-2 cell model, CA exhibited a moderate permeability and was subjected to a mild efflux. Moreover, the apparent permeability coefficient (Papp) of CA transported across Caco-2 cell monolayers was significantly changed when the inhibitors of specific active transporter and passive diffusion were added to cells, suggesting that the absorption and transport of CA involved both passive and active transportation. The present study is an important first step towards understanding the absorption, transport, and metabolic mechanisms of CA. This could provide the scientific basis for developing CA-containing functional foods or dietary supplements with improved bioavailability.

scholarly journals Transepithelial Transport and Metabolism of Boronated Dipeptides Across Caco-2 and HCT-8 Cell Monolayers

1996 ◽  
Vol 3 (6) ◽  
pp. 277-286
Author(s):  
Amy L. Elkins ◽  
John G. Eley ◽  
Merrill C. Miller III ◽  
Iris H. Hall ◽  
Anup Sood ◽  
...  
Keyword(s):  
Oral Delivery ◽  
Cell Monolayer ◽  
Therapeutic Agents ◽  
Transepithelial Transport ◽  
Passive Transport ◽  
Cell Monolayers ◽  
Proteins And Peptides

Oral delivery of proteins and peptides as therapeutic agents is problematic due to their low bioavailability. This study examined the effect of boronation on the transepithelial transport and metabolism of three glycine-phenylalanine dipeptides in Caco-2 and HCT-8 cell monolayers. The three dipeptides exhibited passive transport characteristics in the monolayer systems. However, metabolism of the boronated dipeptides did occur, but to a lesser extent than the non-boronated glycine-phenylalanine dipeptide. The same metabolic scheme was seen in both cell monolayer system, but greater metabolism was seen in the HCT-8 cell monolayers.

scholarly journals Transepithelial Transport Characteristics of the Cholesterol- Lowing Soybean Peptide, WGAPSL, in Caco-2 Cell Monolayers

Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2843
Author(s):  
Huijuan Zhang ◽  
Yawen Duan ◽  
Yulin Feng ◽  
Jing Wang
Keyword(s):  
Tight Junctions ◽  
Soybean Protein ◽  
Blood Stream ◽  
Transepithelial Transport ◽  
Apparent Permeability ◽  
Cholesterol Lowering ◽  
Phase Ii Metabolism ◽  
Cell Monolayers ◽  
Apparent Permeability Coefficient ◽  
Lowering Activity

Recent studies have shown that soybean protein and its peptides have cholesterol-lowering activities. However, it is not clear whether these peptides could overcome physiological barriers, such as phase II metabolism in gastrointestinal tract and poor permeability, to reach the blood stream in its intact form. Therefore, the transepithelial transport characteristics of soybean peptide Trp-Gly-Ala-Pro-Ser-Leu (WGAPSL) with cholesterol- lowering activity were investigated in Caco-2 cells. In this study; the transepithelial absorption of WGAPSL was studied using human intestinal Caco-2 cell monolayers. The results showed that WGAPSL had good stability (83.9% ±1.98%) after simulated gastric and intestinal digestion. During the apical (AP) side to basolateral (BL) side transport, WGAPSL was absorbed intact through Caco-2 cell monolayers with apparent permeability coefficient (Papp) values of 4.4 × 10−8 to 1.2 × 10−8 cm/s. Cytochalasin D loosened the tight junctions of Caco-2 cell monolayers and significantly (p < 0.05) improved the transport process. Sodium azide, wortmannin, and Gly-Pro had minimal effects on transport, demonstrating that the major transport route of WGAPVL was paracellular via tight junctions. Finally, LC-MS analysis showed that Gly-Ala-Pro (GAP) was the important part for the intact absorption of WGAPVL and Trp (W) was the most unstable amino acid residue.

Bioaccessibility in daily diet and bioavailability in vitro of aflatoxins from maize after cooking

2019 ◽  
Vol 12 (2) ◽  
pp. 173-181 ◽  
Author(s):  
X. Lin ◽  
X. Hu ◽  
Y. Zhang ◽  
Y. Xia ◽  
M. Zhang
Keyword(s):  
Cell Model ◽  
Intestinal Transport ◽  
In Vitro Digestion ◽  
Food Type ◽  
Apparent Permeability ◽  
Steam Cooking ◽  
Degradation Rates ◽  
Apparent Permeability Coefficient ◽  
Aflatoxin B

Bioavailability is not a constant percentage of a contaminant in food but is affected by many factors, such as food type, treatment, diet structure and interaction with other compounds. To evaluate these influences, we measured the bioaccessibility of aflatoxins from nine naturally polluted maize samples, collected from southeast China, using an in vitro digestion model, and analysed the intestinal transport of aflatoxins by a Caco-2 cell model. Steam cooking treatment could reduce the aflatoxin levels in maize bread. The degradation rates of aflatoxin B1, aflatoxin B2, aflatoxin G1, and aflatoxin G2 ranged from 24.9±3.2 to 33.9±3.5%, 27.0±2.0 to 39.0±1.8%, 27.9±7.9 to 34.4±8.2% and 25.6±3.6 to 37.2±6.5%, respectively. As a result, the bioaccessibility of aflatoxins determined by an in vitro digestion model (41.5-63.3%) was much lower than the previously reported 80%. Edible oil could increase the bioaccessibility of aflatoxin, whereas lettuce would decrease the exposure amount from maize. With a Caco-2 cell model, the apparent permeability coefficient exceeding 10-5 cm/s indicated that there is high absorption of aflatoxins in the human body, while the intestinal transport can be effectively restrained in the presence of chlorophyll.

scholarly journals Bioavailability Studies of BioTurmin-WD (Water Dispersible Curcuminoids) Using Caco-2 Cell Model

2014 ◽  
Vol 3 (3) ◽  
pp. 158 ◽  
Author(s):  
H. N. Shivaprasad ◽  
M. Bhanumathy ◽  
Subrata Pandit ◽  
D. Manohar ◽  
B. Pavan Kumar ◽  
...  
Keyword(s):  
Cell Model ◽  
Curcuma Longa ◽  
Cell Monolayer ◽  
Absorption Capacity ◽  
Apparent Permeability ◽  
Water Dispersible ◽  
Therapeutic Value ◽  
Clinical Benefits ◽  
Apparent Permeability Coefficient

<p>Curcuminoids is the mixture of curcumin, demethoxycurcumin and bisdemethoxycurcumin obtained from dried rhizomes of <em>Curcuma longa, </em>commonly used for its wide therapeutic value. However, the absorption efficacy of curcuminoids is too low to exhibit its proper therapeutic value. Thus, a new preparation named as BioTurmin-WD (water dispersible curcuminoids) was developed for improved bioavailability. By using human Caco-2 cell monolayer, the permeability efficacy of BioTurmin-WD was evaluated and compared with that of 95% pure curcuminoids. Caco-2 model predicts the<em> in vivo </em>absorption of drugs across the gut wall by measuring the rate of transport of a compound across the Caco-2 cell line. BioTurmin-WD was added to the apical layer and basolateral samples were collected over 120 min to examine the concentration diffusing across the cell monolayer. Permeable curcuminoids across the cell monolayer was analysed through reverse phase high pressure liquid chromatography (RP-HPLC). Apparent permeabilities (P<sub>app</sub>) of BioTurmin-WD and 95% curcuminoids were found to be 5.89 × 10<sup>-6</sup> and 2.65 × 10<sup>-6</sup> cm/s respectively. The apparent permeability coefficient of BioTurmin-WD was 7.03-fold higher than 95% pure curcuminoids. Percentage permeability of BioTurmin-WD (0.2945) was much higher than 95% curcuminoids (0.0859). Results indicated that BioTurmin-WD have a much higher absorption capacity (bioavailability) compared to 95% pure curcuminoids. Thus, BioTurmin-WD may be useful as a dietary supplement with greater bioavailability to exert clinical benefits in humans at a lower dosage.</p>

Processing Immunoperoxidase Labeled Cell Monolayers and Cryostat Sesctions For Electron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 714-715
Author(s):  
K. Chien ◽  
R. Van de Velde ◽  
I.P. Shintaku ◽  
A.F. Sassoon
Keyword(s):  
Cell Monolayer ◽  
Cell Types ◽  
Surface Antigens ◽  
Immunoperoxidase Method ◽  
Reaction Step ◽  
Hot Plate ◽  
Air Drying ◽  
New Approach ◽  
Cell Monolayers ◽  
Glass Slides

Immunoelectron microscopy of neoplastic lymphoma cells is valuable for precise localization of surface antigens and identification of cell types. We have developed a new approach in which the immunohistochemical staining can be evaluated prior to embedding for EM and desired area subsequently selected for ultrathin sectioning.A freshly prepared lymphoma cell suspension is spun onto polylysine hydrobromide- coated glass slides by cytocentrifugation and immediately fixed without air drying in polylysine paraformaldehyde (PLP) fixative. After rinsing in PBS, slides are stained by a 3-step immunoperoxidase method. Cell monolayer is then fixed in buffered 3% glutaraldehyde prior to DAB reaction. After the DAB reaction step, wet monolayers can be examined under LM for presence of brown reaction product and selected monolayers then processed by routine methods for EM and embedded with the Chien Re-embedding Mold. After the polymerization, the epoxy blocks are easily separated from the glass slides by heatingon a 100°C hot plate for 20 seconds.

Improved micro-impedance spectroscopy to determine cell barrier properties

2020 ◽  
Vol 4 (3) ◽  
pp. 150-155 ◽  
Author(s):  
Md. Mehadi Hasan Sohag ◽  
Olivier Nicoud ◽  
Racha Amine ◽  
Abir Khalil-Mgharbel ◽  
Jean-Pierre Alcaraz ◽  
...  
Keyword(s):  
Impedance Spectroscopy ◽  
Epithelial Cells ◽  
Lipid Bilayers ◽  
Cultured Cells ◽  
Cell Model ◽  
Measurement Techniques ◽  
Cell Monolayer ◽  
Microfluidic Channel ◽  
Small Surface ◽  
Resistance Pathway

AbstractThe goal of this study was to determine whether the Tethapod system, which was designed to determine the impedance properties of lipid bilayers, could be used for cell culture in order to utilise micro-impedance spectroscopy to examine further biological applications. To that purpose we have used normal epithelial cells from kidney (RPTEC) and a kidney cancer cell model (786-O). We demonstrate that the Tethapod system is compatible with the culture of 10,000 cells seeded to grow on a small area gold measurement electrode for several days without affecting the cell viability. Furthermore, the range of frequencies for EIS measurements were tuned to examine easily the characteristics of the cell monolayer. We demonstrate significant differences in the paracellular resistance pathway between normal and cancer kidney epithelial cells. Thus, we conclude that this device has advantages for the study of cultured cells that include (i) the configuration of measurement and reference electrodes across a microfluidic channel, and (ii) the small surface area of 6 parallel measurement electrodes (2.1 mm2) integrated in a microfluidic system. These characteristics might improve micro-impedance spectroscopy measurement techniques to provide a simple tool for further studies in the field of the patho-physiology of biological barriers.

Copper repletion enhances apical iron uptake and transepithelial iron transport by Caco-2 cells

2002 ◽  
Vol 282 (3) ◽  
pp. G527-G533 ◽  
Author(s):  
Okhee Han ◽  
Marianne Wessling-Resnick
Keyword(s):  
Iron Uptake ◽  
Iron Transport ◽  
Cell Monolayer ◽  
Transepithelial Transport ◽  
Apical Surface ◽  
Divalent Metal Transporter 1 ◽  
Divalent Metal Transporter ◽  
Metal Transporter ◽  
Copper Status ◽  
Copper Supplementation

The influence of copper status on Caco-2 cell apical iron uptake and transepithelial transport was examined. Cells grown for 7–8 days in media supplemented with 1 μM CuCl2had 10-fold higher cellular levels of copper compared with control. Copper supplementation did not affect the integrity of differentiated Caco-2 cell monolayers grown on microporous membranes. Copper-repleted cells displayed increased uptake of iron as well as increased transport of iron across the cell monolayer. Northern blot analysis revealed that expression of the apical iron transporter divalent metal transporter-1 (DMT1), the basolateral transporter ferroportin-1 (Fpn1), and the putative ferroxidase hephaestin (Heph) was upregulated by copper supplementation, whereas the recently identified ferrireductase duodenal cytochrome b (Dcytb) was not. These results suggest that DMT1, Fpn1, and Heph are involved in the iron uptake process modulated by copper status. Although a clear role for Dcytb was not identified, an apical surface ferrireductase was modulated by copper status, suggesting that its function also contributes to the enhanced iron uptake by copper-repleted cells. A model is proposed wherein copper promotes iron depletion of intestinal Caco-2 cells, creating a deficiency state that induces upregulation of iron transport factors.

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