Activation of Intestinal Cl− Secretion by Lubiprostone Requires the Cystic Fibrosis Transmembrane Conductance Regulator

• Published in: Gastroenterology (New York, N.Y. 1943) Vol. 137; no. 3; pp. 976 - 985
• Main Authors: Bijvelds, Marcel J.C, Bot, Alice G.M, Escher, Johanna C, de Jonge, Hugo R
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-09-2009
• Subjects: Adult; Alprostadil - analogs & derivatives; Alprostadil - pharmacology; Animals; Carbachol - pharmacology; Cell Line, Tumor; Child; Chloride Channels - metabolism; Chlorides - metabolism; Colforsin - pharmacology; Colon - metabolism; Cyclic AMP - metabolism; Cystic Fibrosis Transmembrane Conductance Regulator - metabolism; Dose-Response Relationship, Drug; Gastroenterology and Hepatology; Humans; Ileum - metabolism; Intestinal Mucosa - secretion; Ion Transport; Lubiprostone; Membrane Potentials; Mice; Mice, Knockout; Receptors, Prostaglandin E - antagonists & inhibitors; Receptors, Prostaglandin E - metabolism; Receptors, Prostaglandin E, EP2 Subtype; Tumor Cells, Cultured; CaCC; CF; prostaglandin; Ca 2+-activated Cl − channel; CF transmembrane conductance regulator; IBMX; PKA; PGE 2; isobutyl methylxanthine; short-circuit current; cystic fibrosis; Isc; protein kinase A; CFTR
• ISSN: 0016-5085; 1528-0012
• DOI: 10.1053/j.gastro.2009.05.037

Background & Aims Lubiprostone alleviates constipation by stimulating intestinal fluid secretion, purportedly through activation of ClC-2–type Cl− channels. Intestinal obstruction is also a recurrent cause of distress in cystic fibrosis (CF) patients, caused by loss of CF transmembrane conductance regulator (CFTR) Cl− channel activity. Because ClC-2 recruitment might be beneficial to CF patients, we investigated lubiprostone's mode of action. Methods Cl− transport was measured in an Ussing chamber, in 3 model systems: (1) T84 colonocytes, (2) intestinal epithelium of wild-type and CF mice, and (3) intestinal epithelium of CF patients and controls. Results In T84 monolayers, lubiprostone induced a robust secretory response. Selective permeabilization of the basolateral plasma membrane revealed that lubiprostone activated an apical Cl− conductance. The lubiprostone response was attenuated by H89, an inhibitor of the cAMP-dependent protein kinase, and lubiprostone precluded responsiveness to the cAMP agonist forskolin. CFTR blockage by CFTRinh172, but not ClC-2 blockage by CdCl2 , inhibited the lubiprostone response. Lubiprostone induced a CdCl2 -insensitive secretory response in mouse intestine, but failed to induce intestinal Cl− secretion in Cftr-null mice. Correspondingly, lubiprostone induced a secretory response in human intestinal epithelium, but not in tissue of CF patients. The EP4 -type prostanoid receptor antagonist L-161,982 blocked the lubiprostone response in all 3 models studied. In T84 cells, lubiprostone induced a rise in cAMP levels that was sensitive to EP4 -receptor blockage. Conclusions Lubiprostone enhances intestinal Cl− and fluid secretion via prostanoid receptor signaling, triggering activation of CFTR. Therefore, it is of limited use for treatment of CF-related intestinal disease.