• Journal of Pharmaceutical Investigation
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• v.29 no.4
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• pp.287-293
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• 1999

Transport study of horseradish peroxidase and transferrin-horseradish peroxidase conjugate was performed using an in vitro Caco-2 cell cultured monolayer grown on a polycarbonate membrane of $Transwell^{\circledR}$, Horseradish peroxidase was not transported across Caco-2 cell monolayer. Transferrin-horseradish peroxidase conjugate was transported through Caco-2 cell monolayer. The apparent membrane permeability coefficient $(P_{app})$ of transferrin horseradish peroxidase conjugate was $6.54{\times}10^{-7}\;cm/sec$. The $P_{app}$ value of transferrin-horseradish peroxidase conjugate across Caco-2 cell monolayer was increased to $11.9{\times}10^{-7}\;cm/sec$ in the presence of $50\;{mu}g/ml$ brefeldin-A. These results suggest the transferrin receptor mediated transcytosis of transferrin-horseradish peroxidase conjugate across Caco-2 cell monolayer.

• Food Science of Animal Resources
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• v.26 no.4
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• pp.497-502
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• 2006

The adhesion of 16 bifidobacterial strains, including 10 isolates from Korea infants, to Caco-2 cells and their cell surface hydrophobicity were tested. The results of adhesion and cell surface hydrophobicity of for various bifidobacterial strains were obtained and correlations between adhesion and hydrophobicity were strain-dependent properties. Any correlations between species of tested strains were not observed. Among the tested strains, Bifidobacterim longum D6, B. longum H4, B. thermophilum ATCC 25525, B. suis ATCC 27533, and B. animalis subsp. lactis BB12 had higher adherent properties and B. bifidum B3, B. longum D6, B. longum stronger hydrophobicity, respectively. Due to the strain-dependant correlation between adhesion to Caco-2 cells and cell surface hydrophobicity of bifidobacteria, these results provide a possible method for preliminary selection of bifidobacteria potentially adherent to Caco-2 cells by means of cell surface hydrophobic properties.

• BMB Reports
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• v.32 no.3
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• pp.266-272
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• 1999

The insulin-like growth factor (IGF) system consisting of IGF-I, IGF-II, IGF-receptors, and IGF-binding proteins (IGFBP) regulates the proliferation of a variety of cancer cell types. To examine whether a decrease in endogenous IGFBP-3 stimulates proliferation or inhibits differentiation, Caco-2 cells, a human colon adenocarcinoma cell line, were stably transfected with an anti-sense IGFBP-3 expression construct or pcDNA3 vector as control. Accumulation of IGFBP-3 mRNA and secretion of IGFBP-3 into serum-free conditioned medium, 9 days after plating, were significantly lower in Caco-2 cell clones transfected with anti-sense IGFBP-3 cDNA compared to the controls. The anti-sense clones grew at a similar rate to the controls for 8 days after plating, but achieved a higher final density between days 10 and 12. The levels of sucrase-isomaltase mRNA, a marker of enterocyte differentiation of Caco-2 cells, were lower in the anti-sense clones examined on day 9. In conclusion, proliferation of Caco-2 cells can be stimulated by lowering endogenously-produced IGFBP-3.

• Food Science of Animal Resources
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• v.21 no.2
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• pp.133-141
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• 2001

Adherence of probiotic bacteria to intestinal epithelium is found to be the most principal characteristics among the various physiological functionality. This study was conducted to investigate the effect of bifidobacterial growth properties and condition on the Caco-2 cell adherence and to construct a basic data on adherence-related research. Among 20 strains of bifidobacteris tested, when measured by cell surface hydrophobicity(CSH) and cell agglutination(CA), Bifidobacterium bifidum ATCC29521, Bif. adolescentis K8, and Bif. infantis K9 were selected. Using these strains, variations of Caso-2 cell adherence depending upon experimental condition were analyzed. The results obtained are as follows : Even though Bif. bifidum ATCC29521, Bif. adolescentis K8, and Bif. infantis K9 reached more 85% cell surface hydrophobicity there was no significant difference in cell agglutination, when reached 31.54$\pm$0.54mg/ml. By direct count method for adherence, viable cell count of M3, K1, K2, K8, K9 and K10 reached more 100 counts per 100 Caco-2 cells. When Bif. bifidum ATCC29521, Bif. adolescentistis K8, and Bif. infantis K9 were used to compare the adherence depending upon viable cell counts, reaction time, and growth phase, the more viable cell count, and the more adhered cell counts, the less adherence percentage. In addition, there was no difference in adherence percentage of bifidobacteria when bifidobacteria was incubated from 1 to 8 hrs after Caco-2 cells already formed monolayer. Considering of the effect of growth phase of bifidobacteria on adherence variation, all strains showed the highest adherence during the early stage of stationary phase. In conclusion, adherence of bifidobacteria was affected by strain specificity, viable cell count, and growth activity.

• Natural Product Sciences
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• v.20 no.1
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• pp.1-6
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• 2014

P-glycoprotein (P-gp) is a permeability glycoprotein also known as multidrug resistance protein 1 (MDR1). P-gp is an ATP-binding cassette (ABC) transporter that pumps various types of drugs out of cells. These transporters reduce the intracellular concentrations of drugs and disturb drug absorption. The Caco-2 cell permeability assay system is an effective in vitro system that predicts the intestinal absorption of drugs and the functions of enzymes and transporters. Rhodamine-123 (R-123) and digoxin are well-known P-gp substrates that have been used to determine the function of P-gp. Efflux of P-gp substrates by P-gp has been routinely evaluated. To date, a number of herbal medicines have been tested with Caco-2 cell permeability assay system to assess bioavailability. There are growing efforts to find phytochemicals that potentially regulate P-gp function. The Caco-2 cell permeability assay system is a primary strategy to search for candidates of P-gp inhibitors. In this mini review, we have summarized the P-gp modulation by herbal extracts, decoctions or single components from natural products using Caco-2 cell permeability assays. Many natural products are known to regulate P-gp and herbal medicines could be used in combination with conventional drugs to enhance bioavailability.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.5
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• pp.581-585
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• 2005

Five lactic acid bacteria (LAB) including 2 Lactobacillus strains isolated from Kimchi were evaluated to determine the binding ability to Caco-2 cells and $AFB_1$. LAB were divided into three different groups ; viable, heat-treated, and acid-treated cells. In the radioactive-labeling assay for bound cell counting, viable Lactobacillus Plantarum KCTC 3099 showed the higher adhesion to Caco-2 cells with the binding capacity of $39.2\%$, which was $149\%$ higher than Lactobacillus rhamnosus GG as a positive control. Leuconostoc mesenteroids KCTC 3100 showed the similar binding ability to L. rhamnosus GG. After 1 hour incubation at $37^{\circ}C$ with $AFB_1$, viable L. Planterum KTCC 3099 removed the toxin by $49.8\%$, which was similar level to L. rhamnosus GG. Both heat- and acid-treated groups showed high binding effect but acid-treated group was more effective for both Caco-2 cell binding and $AFB_1$ removal than the other. These results indicate that components of bacterial cell wall might be involved in tile binding to intestinal cells and toxins.

• Parasites, Hosts and Diseases
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• v.52 no.5
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• pp.459-469
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• 2014

Entamoeba histolytica is a tissue-invasive protozoan parasite causing dysentery in humans. During infection of colonic tissues, amoebic trophozoites are able to kill host cells via apoptosis or necrosis, both of which trigger IL-8-mediated acute inflammatory responses. However, the signaling pathways involved in host cell death induced by E. histolytica have not yet been fully defined. In this study, we examined whether calpain plays a role in the cleavage of pro-survival transcription factors during cell death of colonic epithelial cells, induced by live E. histolytica trophozoites. Incubation with amoebic trophozoites induced activation of m-calpain in a time- and dose-dependent manner. Moreover, incubation with amoebae resulted in marked degradation of STAT proteins (STAT3 and STAT5) and NF-${\kappa}B$ (p65) in Caco-2 cells. However, $I{\kappa}B$, an inhibitor of NF-${\kappa}B$, was not cleaved in Caco-2 cells following adherence of E. histolytica. Entamoeba-induced cleavage of STAT proteins and NF-${\kappa}B$ was partially inhibited by pretreatment of cells with a cell-permeable calpain inhibitor, calpeptin. In contrast, E. histolytica did not induce cleavage of caspase-3 in Caco-2 cells. Furthermore, pretreatment of Caco-2 cells with a calpain inhibitor, calpeptin (but not the pan-caspase inhibitor, z-VAD-fmk) or m-calpain siRNA partially reduced Entamoeba-induced DNA fragmentation in Caco-2 cells. These results suggest that calpain plays an important role in E. histolytica-induced degradation of NF-${\kappa}B$ and STATs in colonic epithelial cells, which ultimately accelerates cell death.

• Journal of Applied Biological Chemistry
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• v.63 no.1
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• pp.35-41
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• 2020

(-)-epigallocatechin-3-gallate (EGGG), a flavonoid found in green tea, is known to have low bioavailability. In this study, we determine whether piperine, a natural bioenhancer, can increase the absorption rate of EGCG. Using a Caco-2 cell monolayer, permeability experiments were performed in Hanks' balanced salt solution (HBSS) and EGCG stability was adjusted to pH 6.5 and pH 5.5 by ascorbic acid treatment. When HBSS was adjusted to pH 6.5, EGCG remained at 94.78% for up to 2 h and remained at 86.04% after 4 h and the net efflux decreased compared to the control. As a result, uptake was significantly increased in the piperine co-administered group compared to the EGCG-alone group, showing that piperine increased the permeability of EGCG in the Caco-2 cell monolayer. These results suggest that piperine inhibits EGCG glucuronidation and efflux, allowing for greater absorption of EGCG.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.287-292
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• 2018

Objective: The aim of present study was to assess the anti-oxidant potential of water-soluble peptides (WSPs) extract derived from buffalo and cow milk Cheddar cheeses at different stages of ripening. Methods: The antioxidant potential of WSPs extract was assessed through 2,2'-azinobis-3-ethylbenzothiazoline-6sulfonic acid (ABTS)-radical scavenging activity. In addition, impact of WSPs extract on cell viability and production of reactive oxygen species (ROS) in human colon adenocarcinoma Caco-2 (tert-butylhydroperoxide-induced) cell lines was also evaluated. Results: The ABTS-radical scavenging activity increased progressively with ripening period and dose-dependently in both cheeses. However, peptide extract from buffalo milk Cheddar cheese demonstrated relatively higher activity due to higher contents of water-soluble nitrogen. Intracellular ROS production in Caco-2 cells decreased significantly (p<0.05) till 150th day of cheese ripening and remained constant thereafter. Additionally, dose-dependent response of WSPs extract on antioxidant activity was noticed in the Caco-2 cell line. Conclusion: On the basis of current in vitro study, the Cheddar cheese WSPs extract can protect intestinal epithelium against oxidative stress due to their antioxidant activity.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.343-349
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• 2009

Inhibitory effect of Escherichia coli O157 adhered to Caco-2 cells by the cells of Lactobacillus plantarum K11 and the cell-free culture supernatant (CFCS) and bacteriocin prepared from this strain was investigated. As the cell counts of viable L. plantarum K11 previously adhered to Caco-2 were increased, the rate of adhesion and adherent cell counts of E. coli O157 was lower. However, because the heated L. plantarum K11 rarely have the adhesion ability to Caco-2, the adhesion rate and adherent cell counts of E. coli O157 were high. In addition, the inhibitory effects of E. coli O157 adhesion by the CFCS and bacteriocin of L. plantarum K11 were dose-dependent manner. Therefore, the inhibition of adhesion of E. coli O157 to Caco-2 may result from the antimicrobial substances such as lactic acid and bacteriocin. Moreover the inhibitory activity of adhesion by the heated bacteriocin for 30 min at 100oC was similar to activity of non-treated bacteriocin, but the activity was disappeared by treatment with protease.