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Remodeling of Tight Junctions and Enhancement of Barrier Integrity of the CACO-2 Intestinal Epithelial Cell Layer by Micronutrients.

Valenzano MC, DiGuilio K, Mercado J, Teter M, To J, Ferraro B, Mixson B, Manley I, Baker V, Moore BA, Wertheimer J, Mullin JM - PLoS ONE (2015)

Bottom Line: All five of these chemically very diverse micronutrients increased transepithelial electrical resistance (Rt) significantly, but only berberine also improved barrier integrity to the non-electrolyte D-mannitol.The exact effects of the micronutrients on barrier integrity and TJ protein composition were found to be highly dependent on the degree of differentiation of the cell layer at the time it was exposed to the micronutrient.The substratum to which the epithelial layer adheres was also found to regulate the response of the cell layer to the micronutrient.

View Article: PubMed Central - PubMed

Affiliation: Lankenau Institute for Medical Research, Wynnewood, PA, 19096, United States of America.

ABSTRACT
The micronutrients zinc, quercetin, butyrate, indole and berberine were evaluated for their ability to induce remodeling of epithelial tight junctions (TJs) and enhance barrier integrity in the CACO-2 gastrointestinal epithelial cell culture model. All five of these chemically very diverse micronutrients increased transepithelial electrical resistance (Rt) significantly, but only berberine also improved barrier integrity to the non-electrolyte D-mannitol. Increases of Rt as much as 200% of untreated controls were observed. Each of the five micronutrients also induced unique, signature-like changes in TJ protein composition, suggesting multiple pathways (and TJ arrangements) by which TJ barrier function can be enhanced. Decreases in abundance by as much as 90% were observed for claudin-2, and increases of over 300% could be seen for claudins -5 and -7. The exact effects of the micronutrients on barrier integrity and TJ protein composition were found to be highly dependent on the degree of differentiation of the cell layer at the time it was exposed to the micronutrient. The substratum to which the epithelial layer adheres was also found to regulate the response of the cell layer to the micronutrient. The implications of these findings for therapeutically decreasing morbidity in Inflammatory Bowel Disease are discussed.

No MeSH data available.


Related in: MedlinePlus

The effect of six micronutrients on CACO-2 transepithelial flux of 14C-D-Mannitol.In all cases, after electrical measurements, the same CACO-2 cell layers represented in Fig 1 were used to perform radiotracer flux studies with 0.1mM, 0.20 μCi/ml 14C-D-mannitol, as described in Materials and Methods. Data represents the percent of control flux rate normalized for each experiment, and is expressed as the mean ± standard error for the total number of cell layers per condition, as detailed in Fig 1. * indicates P < 0.05, ** indicates P < 0.01 and *** indicates P < 0.001 (one-way ANOVA followed by Dunnett’s post hoc testing versus control).
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pone.0133926.g002: The effect of six micronutrients on CACO-2 transepithelial flux of 14C-D-Mannitol.In all cases, after electrical measurements, the same CACO-2 cell layers represented in Fig 1 were used to perform radiotracer flux studies with 0.1mM, 0.20 μCi/ml 14C-D-mannitol, as described in Materials and Methods. Data represents the percent of control flux rate normalized for each experiment, and is expressed as the mean ± standard error for the total number of cell layers per condition, as detailed in Fig 1. * indicates P < 0.05, ** indicates P < 0.01 and *** indicates P < 0.001 (one-way ANOVA followed by Dunnett’s post hoc testing versus control).

Mentions: Following the work of other research groups, we had previously shown that a chemically, highly diverse group of micronutrients could exert similar action in enhancing the barrier integrity of an epithelial cell layer [8]. The effect could be to elevate Rt, to decrease Jm or both. Where we had previously shown this to be true for a renal epithelial cell layer, we here show it to be true for the human gastrointestinal (CACO-2) cell layer. Zinc, quercetin, berberine, indole and butyrate all significantly elevated CACO-2 Rt for at least one of the concentrations tested (Fig 1), under conditions described in the figure legend. (The chosen conditions, regarding both micronutrient concentration as well as duration of exposure to micronutrient, tracked reports on the use of these micronutrients in the published literature). Only nicotine failed to affect barrier integrity at any of the concentrations used. In the cases of butyrate and indole, elevations of Rt as much as 100% could be observed. Only berberine, however, was able to simultaneously both increase Rt and decrease Jm significantly, thus evidencing improvement of barrier integrity to two very different solute classes (Fig 2). Considering that these changes in barrier integrity represent changes from the basal state, not changes from a compromised state, the increases in Rt and/or decreases in Jm are quantitatively quite noteworthy.


Remodeling of Tight Junctions and Enhancement of Barrier Integrity of the CACO-2 Intestinal Epithelial Cell Layer by Micronutrients.

Valenzano MC, DiGuilio K, Mercado J, Teter M, To J, Ferraro B, Mixson B, Manley I, Baker V, Moore BA, Wertheimer J, Mullin JM - PLoS ONE (2015)

The effect of six micronutrients on CACO-2 transepithelial flux of 14C-D-Mannitol.In all cases, after electrical measurements, the same CACO-2 cell layers represented in Fig 1 were used to perform radiotracer flux studies with 0.1mM, 0.20 μCi/ml 14C-D-mannitol, as described in Materials and Methods. Data represents the percent of control flux rate normalized for each experiment, and is expressed as the mean ± standard error for the total number of cell layers per condition, as detailed in Fig 1. * indicates P < 0.05, ** indicates P < 0.01 and *** indicates P < 0.001 (one-way ANOVA followed by Dunnett’s post hoc testing versus control).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520484&req=5

pone.0133926.g002: The effect of six micronutrients on CACO-2 transepithelial flux of 14C-D-Mannitol.In all cases, after electrical measurements, the same CACO-2 cell layers represented in Fig 1 were used to perform radiotracer flux studies with 0.1mM, 0.20 μCi/ml 14C-D-mannitol, as described in Materials and Methods. Data represents the percent of control flux rate normalized for each experiment, and is expressed as the mean ± standard error for the total number of cell layers per condition, as detailed in Fig 1. * indicates P < 0.05, ** indicates P < 0.01 and *** indicates P < 0.001 (one-way ANOVA followed by Dunnett’s post hoc testing versus control).
Mentions: Following the work of other research groups, we had previously shown that a chemically, highly diverse group of micronutrients could exert similar action in enhancing the barrier integrity of an epithelial cell layer [8]. The effect could be to elevate Rt, to decrease Jm or both. Where we had previously shown this to be true for a renal epithelial cell layer, we here show it to be true for the human gastrointestinal (CACO-2) cell layer. Zinc, quercetin, berberine, indole and butyrate all significantly elevated CACO-2 Rt for at least one of the concentrations tested (Fig 1), under conditions described in the figure legend. (The chosen conditions, regarding both micronutrient concentration as well as duration of exposure to micronutrient, tracked reports on the use of these micronutrients in the published literature). Only nicotine failed to affect barrier integrity at any of the concentrations used. In the cases of butyrate and indole, elevations of Rt as much as 100% could be observed. Only berberine, however, was able to simultaneously both increase Rt and decrease Jm significantly, thus evidencing improvement of barrier integrity to two very different solute classes (Fig 2). Considering that these changes in barrier integrity represent changes from the basal state, not changes from a compromised state, the increases in Rt and/or decreases in Jm are quantitatively quite noteworthy.

Bottom Line: All five of these chemically very diverse micronutrients increased transepithelial electrical resistance (Rt) significantly, but only berberine also improved barrier integrity to the non-electrolyte D-mannitol.The exact effects of the micronutrients on barrier integrity and TJ protein composition were found to be highly dependent on the degree of differentiation of the cell layer at the time it was exposed to the micronutrient.The substratum to which the epithelial layer adheres was also found to regulate the response of the cell layer to the micronutrient.

View Article: PubMed Central - PubMed

Affiliation: Lankenau Institute for Medical Research, Wynnewood, PA, 19096, United States of America.

ABSTRACT
The micronutrients zinc, quercetin, butyrate, indole and berberine were evaluated for their ability to induce remodeling of epithelial tight junctions (TJs) and enhance barrier integrity in the CACO-2 gastrointestinal epithelial cell culture model. All five of these chemically very diverse micronutrients increased transepithelial electrical resistance (Rt) significantly, but only berberine also improved barrier integrity to the non-electrolyte D-mannitol. Increases of Rt as much as 200% of untreated controls were observed. Each of the five micronutrients also induced unique, signature-like changes in TJ protein composition, suggesting multiple pathways (and TJ arrangements) by which TJ barrier function can be enhanced. Decreases in abundance by as much as 90% were observed for claudin-2, and increases of over 300% could be seen for claudins -5 and -7. The exact effects of the micronutrients on barrier integrity and TJ protein composition were found to be highly dependent on the degree of differentiation of the cell layer at the time it was exposed to the micronutrient. The substratum to which the epithelial layer adheres was also found to regulate the response of the cell layer to the micronutrient. The implications of these findings for therapeutically decreasing morbidity in Inflammatory Bowel Disease are discussed.

No MeSH data available.


Related in: MedlinePlus