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Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities

Secretory diarrhea, which primarily originates through intestinal pathogens and viruses, is a health burden in many regions worldwide. Enterocyte Cl(−) channels, as the final step in enterotoxin-induced fluid secretion, constitute an attractive class of targets for diarrhea therapy. Chloride channel...

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Autores principales: Yu, Bo, Zhu, Xiaojuan, Yang, Xinyu, Jin, Lingling, Xu, Jia, Ma, Tonghui, Yang, Hong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6795057/
https://www.ncbi.nlm.nih.gov/pubmed/31649543
http://dx.doi.org/10.3389/fphar.2019.01181
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author Yu, Bo
Zhu, Xiaojuan
Yang, Xinyu
Jin, Lingling
Xu, Jia
Ma, Tonghui
Yang, Hong
author_facet Yu, Bo
Zhu, Xiaojuan
Yang, Xinyu
Jin, Lingling
Xu, Jia
Ma, Tonghui
Yang, Hong
author_sort Yu, Bo
collection PubMed
description Secretory diarrhea, which primarily originates through intestinal pathogens and viruses, is a health burden in many regions worldwide. Enterocyte Cl(−) channels, as the final step in enterotoxin-induced fluid secretion, constitute an attractive class of targets for diarrhea therapy. Chloride channel inhibitors have become a new class of candidates for antisecretion and anti-intestinal motility agents. In the present study, we identified plumbagin as a transmembrane protein 16A (TMEM16A) inhibitor in a cell-based fluorescence-quenching assay, and the IC(50) value was ∼12.46 µM. Short-circuit current measurements showed that plumbagin reversibly inhibited the E(act)-induced Cl(−) current on the apical side of TMEM16A-transfected Fischer rat thyroid (FRT) cells with no significant effect on cytoplasmic Ca(2+) signaling. Notably, plumbagin also inhibited the activity of intestinal epithelial calcium-activated chloride channel (CaCC) and cystic fibrosis transmembrane conductance regulator (CFTR) in both HT-29 cells and mouse colons, but had no effects on the activity of the Na(+)-K(+) ATPase or K(+) channels. In in vivo experiments, the administration of plumbagin reduced both Escherichia coli heat-stable enterotoxin (STa)- and cholera toxin (CT)-induced intestinal fluid secretion. In neonatal mouse models of CT- and rotavirus infection-induced diarrhea, 0.4 µg plumbagin inhibited secretory diarrhea by >40% and 50%, respectively, without affecting intestinal epithelial integrity or the rotaviral infection. In addition, plumbagin exerted inhibitory effects on the vasoactive intestinal peptide (VIP)-, prostaglandin E2 (PGE2)-, and 5-hydroxytryptamine (5-HT)-stimulated Cl(−) currents. In the evaluations of intestinal motility, plumbagin significantly delayed intestinal motility and inhibited intestinal smooth muscle contractility without an evident impact on contractive frequency. Collectively, our results indicate that plumbagin inhibits both Ca(2+)- and cAMP-activated Cl(−) channels, accounting for the mechanisms of plumbagin inhibition of chloride secretion and intestinal motility. Thus, plumbagin can be a lead compound in the treatment of CT-induced, Traveler’s, and rotaviral diarrhea, as well as other types of secretory diarrhea that result from excessive intestinal fluid secretion and increased intestinal peristalsis.
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spelling pubmed-67950572019-10-24 Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities Yu, Bo Zhu, Xiaojuan Yang, Xinyu Jin, Lingling Xu, Jia Ma, Tonghui Yang, Hong Front Pharmacol Pharmacology Secretory diarrhea, which primarily originates through intestinal pathogens and viruses, is a health burden in many regions worldwide. Enterocyte Cl(−) channels, as the final step in enterotoxin-induced fluid secretion, constitute an attractive class of targets for diarrhea therapy. Chloride channel inhibitors have become a new class of candidates for antisecretion and anti-intestinal motility agents. In the present study, we identified plumbagin as a transmembrane protein 16A (TMEM16A) inhibitor in a cell-based fluorescence-quenching assay, and the IC(50) value was ∼12.46 µM. Short-circuit current measurements showed that plumbagin reversibly inhibited the E(act)-induced Cl(−) current on the apical side of TMEM16A-transfected Fischer rat thyroid (FRT) cells with no significant effect on cytoplasmic Ca(2+) signaling. Notably, plumbagin also inhibited the activity of intestinal epithelial calcium-activated chloride channel (CaCC) and cystic fibrosis transmembrane conductance regulator (CFTR) in both HT-29 cells and mouse colons, but had no effects on the activity of the Na(+)-K(+) ATPase or K(+) channels. In in vivo experiments, the administration of plumbagin reduced both Escherichia coli heat-stable enterotoxin (STa)- and cholera toxin (CT)-induced intestinal fluid secretion. In neonatal mouse models of CT- and rotavirus infection-induced diarrhea, 0.4 µg plumbagin inhibited secretory diarrhea by >40% and 50%, respectively, without affecting intestinal epithelial integrity or the rotaviral infection. In addition, plumbagin exerted inhibitory effects on the vasoactive intestinal peptide (VIP)-, prostaglandin E2 (PGE2)-, and 5-hydroxytryptamine (5-HT)-stimulated Cl(−) currents. In the evaluations of intestinal motility, plumbagin significantly delayed intestinal motility and inhibited intestinal smooth muscle contractility without an evident impact on contractive frequency. Collectively, our results indicate that plumbagin inhibits both Ca(2+)- and cAMP-activated Cl(−) channels, accounting for the mechanisms of plumbagin inhibition of chloride secretion and intestinal motility. Thus, plumbagin can be a lead compound in the treatment of CT-induced, Traveler’s, and rotaviral diarrhea, as well as other types of secretory diarrhea that result from excessive intestinal fluid secretion and increased intestinal peristalsis. Frontiers Media S.A. 2019-10-09 /pmc/articles/PMC6795057/ /pubmed/31649543 http://dx.doi.org/10.3389/fphar.2019.01181 Text en Copyright © 2019 Yu, Zhu, Yang, Jin, Xu, Ma and Yang http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Yu, Bo
Zhu, Xiaojuan
Yang, Xinyu
Jin, Lingling
Xu, Jia
Ma, Tonghui
Yang, Hong
Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities
title Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities
title_full Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities
title_fullStr Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities
title_full_unstemmed Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities
title_short Plumbagin Prevents Secretory Diarrhea by Inhibiting CaCC and CFTR Channel Activities
title_sort plumbagin prevents secretory diarrhea by inhibiting cacc and cftr channel activities
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6795057/
https://www.ncbi.nlm.nih.gov/pubmed/31649543
http://dx.doi.org/10.3389/fphar.2019.01181
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