Eun-A Ko1, Byung-Ju Jin1, Wan Namkung2, Tonghui Ma3, Jay R Thiagarajah4, A S Verkman1. 1. Departments of Medicine and Physiology, University of California, San Francisco, California, USA. 2. College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University, Incheon, Korea. 3. Department of Physiology, Dalian Medical University, Dalian, China. 4. Department of Gastroenterology, Hepatology and Nutrition, Children's Hospital, Boston, Harvard Medical School, Boston, Massachusetts, USA.
Abstract
BACKGROUND: Rotavirus is the most common cause of severe secretory diarrhoea in infants and young children globally. The rotaviral enterotoxin, NSP4, has been proposed to stimulate calcium-activated chloride channels (CaCC) on the apical plasma membrane of intestinal epithelial cells. We previously identified red wine and small molecule CaCC inhibitors. OBJECTIVE: To investigate the efficacy of a red wine extract and a synthetic small molecule, CaCCinh-A01, in inhibiting intestinal CaCCs and rotaviral diarrhoea. DESIGN: Inhibition of CaCC-dependent current was measured in T84 cells and mouse ileum. The effectiveness of an orally administered wine extract and CaCCinh-A01 in inhibiting diarrhoea in vivo was determined in a neonatal mouse model of rotaviral infection. RESULTS: Screening of ∼150 red wines revealed a Cabernet Sauvignon that inhibited CaCC current in T84 cells with IC50 at a ∼1:200 dilution, and higher concentrations producing 100% inhibition. A >1 kdalton wine extract prepared by dialysis, which retained full inhibition activity, blocked CaCC current in T84 cells and mouse intestine. In rotavirus-inoculated mice, oral administration of the wine extract prevented diarrhoea by inhibition of intestinal fluid secretion without affecting rotaviral infection. The wine extract did not inhibit the cystic fibrosis chloride channel (CFTR) in cell cultures, nor did it prevent watery stools in neonatal mice administered cholera toxin, which activates CFTR-dependent fluid secretion. CaCCinh-A01 also inhibited rotaviral diarrhoea. CONCLUSIONS: Our results support a pathogenic role for enterocyte CaCCs in rotaviral diarrhoea and demonstrate the antidiarrhoeal action of CaCC inhibition by an alcohol-free, red wine extract and by a synthetic small molecule. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
BACKGROUND: Rotavirus is the most common cause of severe secretory diarrhoea in infants and young children globally. The rotaviral enterotoxin, NSP4, has been proposed to stimulate calcium-activated chloride channels (CaCC) on the apical plasma membrane of intestinal epithelial cells. We previously identified red wine and small molecule CaCC inhibitors. OBJECTIVE: To investigate the efficacy of a red wine extract and a synthetic small molecule, CaCCinh-A01, in inhibiting intestinal CaCCs and rotaviral diarrhoea. DESIGN: Inhibition of CaCC-dependent current was measured in T84 cells and mouse ileum. The effectiveness of an orally administered wine extract and CaCCinh-A01 in inhibiting diarrhoea in vivo was determined in a neonatal mouse model of rotaviral infection. RESULTS: Screening of ∼150 red wines revealed a Cabernet Sauvignon that inhibited CaCC current in T84 cells with IC50 at a ∼1:200 dilution, and higher concentrations producing 100% inhibition. A >1 kdalton wine extract prepared by dialysis, which retained full inhibition activity, blocked CaCC current in T84 cells and mouse intestine. In rotavirus-inoculatedmice, oral administration of the wine extract prevented diarrhoea by inhibition of intestinal fluid secretion without affecting rotaviral infection. The wine extract did not inhibit the cystic fibrosischloride channel (CFTR) in cell cultures, nor did it prevent watery stools in neonatal mice administered cholera toxin, which activates CFTR-dependent fluid secretion. CaCCinh-A01 also inhibited rotaviral diarrhoea. CONCLUSIONS: Our results support a pathogenic role for enterocyte CaCCs in rotaviral diarrhoea and demonstrate the antidiarrhoeal action of CaCC inhibition by an alcohol-free, red wine extract and by a synthetic small molecule. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
Entities:
Keywords:
Diarrhoea; Intestinal Secretion; Ion Channels
Authors: L J Galietta; M F Springsteel; M Eda; E J Niedzinski; K By; M J Haddadin; M J Kurth; M H Nantz; A S Verkman Journal: J Biol Chem Date: 2001-03-21 Impact factor: 5.157
Authors: Paul A Rufo; Patricia W Lin; Adriana Andrade; Lianwei Jiang; Lucia Rameh; Charles Flexner; Seth L Alper; Wayne I Lencer Journal: Am J Physiol Cell Physiol Date: 2003-12-30 Impact factor: 4.249
Authors: M P Osborne; S J Haddon; A J Spencer; J Collins; W G Starkey; T S Wallis; G J Clarke; K J Worton; D C Candy; J Stephen Journal: J Pediatr Gastroenterol Nutr Date: 1988 Mar-Apr Impact factor: 2.839
Authors: Onur Cil; Puay-Wah Phuan; Anne Marie Gillespie; Sujin Lee; Lukmanee Tradtrantip; Jianyi Yin; Ming Tse; Nicholas C Zachos; Ruxian Lin; Mark Donowitz; Alan S Verkman Journal: FASEB J Date: 2016-11-08 Impact factor: 5.191
Authors: Jay R Thiagarajah; Eun-A Ko; Lukmanee Tradtrantip; Mark Donowitz; A S Verkman Journal: Clin Gastroenterol Hepatol Date: 2013-12-04 Impact factor: 11.382
Authors: Sue E Crawford; Sasirekha Ramani; Jacqueline E Tate; Umesh D Parashar; Lennart Svensson; Marie Hagbom; Manuel A Franco; Harry B Greenberg; Miguel O'Ryan; Gagandeep Kang; Ulrich Desselberger; Mary K Estes Journal: Nat Rev Dis Primers Date: 2017-11-09 Impact factor: 52.329