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Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer

Vasoactive intestinal peptide (VIP) as a neurocrine factor released by enteric neurons has been postulated to participate in the regulation of transcellular active calcium transport across intestinal epithelium, but the preceding evidence is scant and inconclusive. Herein, transepithelial calcium fl...

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Autores principales: Rodrat, Mayuree, Wongdee, Kannikar, Teerapornpuntakit, Jarinthorn, Thongbunchoo, Jirawan, Tanramluk, Duangrudee, Aeimlapa, Ratchaneevan, Thammayon, Nithipak, Thonapan, Natchayaporn, Wattano, Pathnaree, Charoenphandhu, Narattaphol
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9674163/
https://www.ncbi.nlm.nih.gov/pubmed/36399482
http://dx.doi.org/10.1371/journal.pone.0277096
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author Rodrat, Mayuree
Wongdee, Kannikar
Teerapornpuntakit, Jarinthorn
Thongbunchoo, Jirawan
Tanramluk, Duangrudee
Aeimlapa, Ratchaneevan
Thammayon, Nithipak
Thonapan, Natchayaporn
Wattano, Pathnaree
Charoenphandhu, Narattaphol
author_facet Rodrat, Mayuree
Wongdee, Kannikar
Teerapornpuntakit, Jarinthorn
Thongbunchoo, Jirawan
Tanramluk, Duangrudee
Aeimlapa, Ratchaneevan
Thammayon, Nithipak
Thonapan, Natchayaporn
Wattano, Pathnaree
Charoenphandhu, Narattaphol
author_sort Rodrat, Mayuree
collection PubMed
description Vasoactive intestinal peptide (VIP) as a neurocrine factor released by enteric neurons has been postulated to participate in the regulation of transcellular active calcium transport across intestinal epithelium, but the preceding evidence is scant and inconclusive. Herein, transepithelial calcium flux and epithelial electrical parameters were determined by Ussing chamber technique with radioactive tracer in the intestinal epithelium-like Caco-2 monolayer grown on Snapwell. After 3-day culture, Caco-2 cells expressed mRNA of calcium transporters, i.e., TRPV6, calbindin-D(9k), PMCA(1b) and NCX1, and exhibited transepithelial resistance of ~200 Ω cm(2), a characteristic of leaky epithelium similar to the small intestine. VIP receptor agonist was able to enhance transcellular calcium flux, whereas VIP receptor antagonist totally abolished calcium fluxes induced by 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. Since the intestinal cystic fibrosis transmembrane conductance regulator (CFTR) could be activated by VIP and calciotropic hormones, particularly parathyroid hormone, we sought to determine whether CFTR also contributed to the 1,25(OH)(2)D(3)-induced calcium transport. A selective CFTR inhibitor (20–200 μM CFTR(inh)-172) appeared to diminish calcium fluxes as well as transepithelial potential difference and short-circuit current, both of which indicated a decrease in electrogenic ion transport. On the other hand, 50 μM genistein—a molecule that could rapidly activate CFTR—was found to increase calcium transport. Our in silico molecular docking analysis confirmed direct binding of CFTR(inh)-172 and genistein to CFTR channels. In conclusion, VIP and CFTR apparently contributed to the intestinal calcium transport, especially in the presence of 1,25(OH)(2)D(3), thereby supporting the existence of the neurocrine control of intestinal calcium absorption.
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spelling pubmed-96741632022-11-19 Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer Rodrat, Mayuree Wongdee, Kannikar Teerapornpuntakit, Jarinthorn Thongbunchoo, Jirawan Tanramluk, Duangrudee Aeimlapa, Ratchaneevan Thammayon, Nithipak Thonapan, Natchayaporn Wattano, Pathnaree Charoenphandhu, Narattaphol PLoS One Research Article Vasoactive intestinal peptide (VIP) as a neurocrine factor released by enteric neurons has been postulated to participate in the regulation of transcellular active calcium transport across intestinal epithelium, but the preceding evidence is scant and inconclusive. Herein, transepithelial calcium flux and epithelial electrical parameters were determined by Ussing chamber technique with radioactive tracer in the intestinal epithelium-like Caco-2 monolayer grown on Snapwell. After 3-day culture, Caco-2 cells expressed mRNA of calcium transporters, i.e., TRPV6, calbindin-D(9k), PMCA(1b) and NCX1, and exhibited transepithelial resistance of ~200 Ω cm(2), a characteristic of leaky epithelium similar to the small intestine. VIP receptor agonist was able to enhance transcellular calcium flux, whereas VIP receptor antagonist totally abolished calcium fluxes induced by 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. Since the intestinal cystic fibrosis transmembrane conductance regulator (CFTR) could be activated by VIP and calciotropic hormones, particularly parathyroid hormone, we sought to determine whether CFTR also contributed to the 1,25(OH)(2)D(3)-induced calcium transport. A selective CFTR inhibitor (20–200 μM CFTR(inh)-172) appeared to diminish calcium fluxes as well as transepithelial potential difference and short-circuit current, both of which indicated a decrease in electrogenic ion transport. On the other hand, 50 μM genistein—a molecule that could rapidly activate CFTR—was found to increase calcium transport. Our in silico molecular docking analysis confirmed direct binding of CFTR(inh)-172 and genistein to CFTR channels. In conclusion, VIP and CFTR apparently contributed to the intestinal calcium transport, especially in the presence of 1,25(OH)(2)D(3), thereby supporting the existence of the neurocrine control of intestinal calcium absorption. Public Library of Science 2022-11-18 /pmc/articles/PMC9674163/ /pubmed/36399482 http://dx.doi.org/10.1371/journal.pone.0277096 Text en © 2022 Rodrat et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Rodrat, Mayuree
Wongdee, Kannikar
Teerapornpuntakit, Jarinthorn
Thongbunchoo, Jirawan
Tanramluk, Duangrudee
Aeimlapa, Ratchaneevan
Thammayon, Nithipak
Thonapan, Natchayaporn
Wattano, Pathnaree
Charoenphandhu, Narattaphol
Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer
title Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer
title_full Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer
title_fullStr Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer
title_full_unstemmed Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer
title_short Vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like Caco-2 monolayer
title_sort vasoactive intestinal peptide and cystic fibrosis transmembrane conductance regulator contribute to the transepithelial calcium transport across intestinal epithelium-like caco-2 monolayer
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9674163/
https://www.ncbi.nlm.nih.gov/pubmed/36399482
http://dx.doi.org/10.1371/journal.pone.0277096
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