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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 the cell layer substratum on CACO-2 epithelial barrier’s response to zinc CACO-2 cell layers were seeded at identical densities on Millicell PCF and Millicell HA units as described in Materials and Methods.After 7 days, cell layers were refed in apical and basal-lateral compartments with control medium or medium containing 100μM zinc, 48 hrs before electrical measurements. Data represents the percentage of normalized control resistance, normalized control short circuit current, and normalized control flux rate (each relative to proper control; 2 experiments, 4 cell layers per condition). Data shown is expressed as the mean ± standard error of 8 cell layers per condition. P values (Student’s t test, two-tailed) are indicated for statistical comparisons of the various conditions.
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pone.0133926.g007: The effect of the cell layer substratum on CACO-2 epithelial barrier’s response to zinc CACO-2 cell layers were seeded at identical densities on Millicell PCF and Millicell HA units as described in Materials and Methods.After 7 days, cell layers were refed in apical and basal-lateral compartments with control medium or medium containing 100μM zinc, 48 hrs before electrical measurements. Data represents the percentage of normalized control resistance, normalized control short circuit current, and normalized control flux rate (each relative to proper control; 2 experiments, 4 cell layers per condition). Data shown is expressed as the mean ± standard error of 8 cell layers per condition. P values (Student’s t test, two-tailed) are indicated for statistical comparisons of the various conditions.

Mentions: For the purposes of this study however, it was more pertinent that the response of the cell layers to supplemental zinc was quite different, depending on the filter used. Cell layers on HA filters showed a much smaller Rt increase in response to zinc treatment, and actually increased their permeability to mannitol, whereas those on PCF filters exhibited a much greater Rt increase and did not increase their permeability to mannitol (Fig 7). More pointedly, cell layers on HA filters decreased their Isc by almost 20% in response to supplemental zinc, whereas cell layers on PCF filters increased their Isc by over 20% in response to supplemental zinc. Along with degree of differentiation, epithelial substratum thus appears to also play a likely regulatory effect on the action of micronutrients with regard to epithelial barrier properties.


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 the cell layer substratum on CACO-2 epithelial barrier’s response to zinc CACO-2 cell layers were seeded at identical densities on Millicell PCF and Millicell HA units as described in Materials and Methods.After 7 days, cell layers were refed in apical and basal-lateral compartments with control medium or medium containing 100μM zinc, 48 hrs before electrical measurements. Data represents the percentage of normalized control resistance, normalized control short circuit current, and normalized control flux rate (each relative to proper control; 2 experiments, 4 cell layers per condition). Data shown is expressed as the mean ± standard error of 8 cell layers per condition. P values (Student’s t test, two-tailed) are indicated for statistical comparisons of the various conditions.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0133926.g007: The effect of the cell layer substratum on CACO-2 epithelial barrier’s response to zinc CACO-2 cell layers were seeded at identical densities on Millicell PCF and Millicell HA units as described in Materials and Methods.After 7 days, cell layers were refed in apical and basal-lateral compartments with control medium or medium containing 100μM zinc, 48 hrs before electrical measurements. Data represents the percentage of normalized control resistance, normalized control short circuit current, and normalized control flux rate (each relative to proper control; 2 experiments, 4 cell layers per condition). Data shown is expressed as the mean ± standard error of 8 cell layers per condition. P values (Student’s t test, two-tailed) are indicated for statistical comparisons of the various conditions.
Mentions: For the purposes of this study however, it was more pertinent that the response of the cell layers to supplemental zinc was quite different, depending on the filter used. Cell layers on HA filters showed a much smaller Rt increase in response to zinc treatment, and actually increased their permeability to mannitol, whereas those on PCF filters exhibited a much greater Rt increase and did not increase their permeability to mannitol (Fig 7). More pointedly, cell layers on HA filters decreased their Isc by almost 20% in response to supplemental zinc, whereas cell layers on PCF filters increased their Isc by over 20% in response to supplemental zinc. Along with degree of differentiation, epithelial substratum thus appears to also play a likely regulatory effect on the action of micronutrients with regard to epithelial barrier properties.

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