{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T03:23:27Z","timestamp":1785813807314,"version":"3.56.0"},"reference-count":48,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2025,8,26]],"date-time":"2025-08-26T00:00:00Z","timestamp":1756166400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100016263","name":"University Research Board, American University of Beirut","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100016263","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>The Na\u2009\u207a\u2009\/K\u2009\u207a\u2009ATPase, commonly known as the Na\u2009\u207a\u2009\/K\u207a pump, plays a crucial role in colonic sodium and water transport, inducing diarrhea or constipation. Inflammatory bowel disease is often associated with diarrhea and elevated levels of sphingosine-1-phosphate (S1P), suggesting a potential relationship between the pump and S1P. This study investigated the effects of S1P on colonic Na\u2009\u207a\u2009\/K\u2009\u207a\u2009ATPase using Caco-2 cells as a model and the S1P analogue used in the treatment of multiple sclerosis, FTY720P. The pump activity was assessed by measuring the amount of inorganic phosphate released in the presence and absence of ouabain, an ATPase inhibitor. FTY720P induced an inhibition of Na\u2009\u207a\u2009\/K\u2009\u207a\u2009ATPase at 15 minutes that was studied in a previous work. This inhibition shifted however, to stimulation at 2 hours, an effect that was abolished in the presence of JTE-013, an S1PR2 antagonist, and was replicated by CYM5520, an S1PR2 agonist. Further mechanistic exploration revealed that when PKC, NF-\u03baB, COX, PKA, and PI3K enzymes were inhibited, FTY720P no longer influenced Na\u2009\u207a\u2009\/K\u2009\u207a\u2009ATPase activity, indicating their involvement in the signaling cascade. Additional evidence supporting this pathway came from activators of these kinases and exogenous PGE\u2082, both of which stimulated the pump. The results indicate that FTY720P stimulates Na\u2009\u207a\u2009\/K\u2009\u207a\u2009ATPase at 2 hours by binding to S1PR2, leading to PKC activation, followed by NF-\u03baB-mediated induction of PGE\u2082 synthesis. PGE\u2082 then binds to its EP4 receptors, activating PKA and PI3K, ultimately resulting in an increase in the pump\u2019s activity. These findings will open the door to targeted regulation of these intermediate molecules which could potentially alleviate certain undesirable effects of the drug.<\/jats:p>","DOI":"10.1371\/journal.pone.0330818","type":"journal-article","created":{"date-parts":[[2025,8,26]],"date-time":"2025-08-26T17:26:46Z","timestamp":1756229206000},"page":"e0330818","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Mediators of the stimulatory effect of S1P on colonic Na+\/K+ ATPase"],"prefix":"10.1371","volume":"20","author":[{"given":"Maysoon","family":"Noureddine","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4149-4081","authenticated-orcid":true,"given":"Sawsan","family":"Kreydiyyeh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,8,26]]},"reference":[{"issue":"9","key":"pone.0330818.ref001","doi-asserted-by":"crossref","first-page":"2211","DOI":"10.1016\/j.cgh.2022.06.030","article-title":"Global Hospitalization Trends for Crohn\u2019s Disease and Ulcerative Colitis in the 21st Century: A Systematic Review With Temporal Analyses","volume":"21","author":"MJ Buie","year":"2023","journal-title":"Clin Gastroenterol Hepatol"},{"issue":"5","key":"pone.0330818.ref002","first-page":"527","article-title":"Inflammatory bowel disease burden in the Middle East and North Africa Region: a comprehensive analysis of incidence, prevalence, and mortality from 1990-2019","volume":"37","author":"S Alsakarneh","year":"2024","journal-title":"Ann Gastroenterol"},{"issue":"12","key":"pone.0330818.ref003","doi-asserted-by":"crossref","first-page":"2926","DOI":"10.1097\/MIB.0000000000000504","article-title":"Mechanisms Underlying Dysregulation of Electrolyte Absorption in Inflammatory Bowel Disease-Associated Diarrhea","volume":"21","author":"S Priyamvada","year":"2015","journal-title":"Inflamm Bowel Dis"},{"issue":"4","key":"pone.0330818.ref004","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1007\/BF01536025","article-title":"Inverse relationship between colonic (Na+ + K+)-ATPase activity and degree of mucosal inflammation in inflammatory bowel disease","volume":"33","author":"H Allgayer","year":"1988","journal-title":"Dig Dis Sci"},{"issue":"5","key":"pone.0330818.ref005","first-page":"469","article-title":"Sphingosine-1-phosphate in inflammatory bowel disease and colitis-associated colon cancer: the fat\u2019s in the fire","volume":"4","author":"JH Suh","year":"2015","journal-title":"Transl Cancer Res"},{"key":"pone.0330818.ref006","doi-asserted-by":"crossref","first-page":"1235656","DOI":"10.3389\/fphys.2023.1235656","article-title":"The role of sphingosine-1-phosphate in the gut mucosal microenvironment and inflammatory bowel diseases","volume":"14","author":"F Zou","year":"2023","journal-title":"Front Physiol"},{"issue":"1","key":"pone.0330818.ref007","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/1476-9255-3-5","article-title":"Changes in ion transport in inflammatory disease","volume":"3","author":"M Eisenhut","year":"2006","journal-title":"J Inflamm"},{"issue":"7","key":"pone.0330818.ref008","doi-asserted-by":"crossref","DOI":"10.1152\/ajpgi.00369.2015","article-title":"Role of epithelial ion transports in inflammatory bowel disease","volume":"310","author":"D Magalh\u00e3es","year":"2016","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"issue":"3","key":"pone.0330818.ref009","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1007\/s12026-015-8625-5","article-title":"SEW2871 protects from experimental colitis through reduced epithelial cell apoptosis and improved barrier function in interleukin-10 gene-deficient mice","volume":"61","author":"J Dong","year":"2015","journal-title":"Immunol Res"},{"issue":"6","key":"pone.0330818.ref010","doi-asserted-by":"crossref","first-page":"2077","DOI":"10.1002\/jcb.24079","article-title":"Sphingosine-1-phosphate is a mediator of TNF-\u03b1 action on the Na+\/K+ ATPase in HepG2 cells","volume":"113","author":"Z Dakroub","year":"2012","journal-title":"J Cell Biochem"},{"key":"pone.0330818.ref011","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.lfs.2018.11.026","article-title":"FTY720P inhibits the Na+\/K+ ATPase in Caco-2 cells via S1PR2: PGE2 and NO are along the signaling pathway","volume":"215","author":"R Rida","year":"2018","journal-title":"Life Sci"},{"key":"pone.0330818.ref012","unstructured":"U.S. Food and Drug Administration. Highlights of prescribing information: fingolimod. 2016. https:\/\/www.accessdata.fda.gov\/drugsatfda_docs\/label\/2016\/022527s018lbl.pdf"},{"issue":"2","key":"pone.0330818.ref013","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0193139","article-title":"Epinephrine modulates Na+\/K+ ATPase activity in Caco-2 cells via Src, p38MAPK, ERK and PGE2","volume":"13","author":"L El Moussawi","year":"2018","journal-title":"PLoS ONE"},{"key":"pone.0330818.ref014","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/0076-6879(88)56013-5","article-title":"ATPase and phosphatase activity of Na+,K+-ATPase: molar and specific activity, protein determination","volume":"156","author":"M Esmann","year":"1988","journal-title":"Methods Enzymol"},{"issue":"2","key":"pone.0330818.ref015","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1016\/S0021-9258(18)66180-0","article-title":"A microcolorimetric method for the determination of inorganic 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Hla","year":"2001","journal-title":"Science"},{"issue":"19","key":"pone.0330818.ref021","doi-asserted-by":"crossref","first-page":"3850","DOI":"10.1016\/j.jmb.2016.08.002","article-title":"Regulation, Signaling, and Physiological Functions of G-Proteins","volume":"428","author":"V Syrovatkina","year":"2016","journal-title":"J Mol Biol"},{"issue":"1","key":"pone.0330818.ref022","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1159\/000186689","article-title":"Functions and regulatory mechanisms of Gq-signaling pathways","volume":"17","author":"N Mizuno","year":"2009","journal-title":"Neurosignals"},{"issue":"5573","key":"pone.0330818.ref023","doi-asserted-by":"crossref","first-page":"1636","DOI":"10.1126\/science.1071550","article-title":"G protein pathways","volume":"296","author":"SR Neves","year":"2002","journal-title":"Science"},{"issue":"1","key":"pone.0330818.ref024","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1091\/mbc.11.1.39","article-title":"Is phosphorylation of the alpha1 subunit at Ser-16 involved in the control of Na,K-ATPase activity by phorbol ester-activated protein kinase C?","volume":"11","author":"E F\u00e9raille","year":"2000","journal-title":"Mol Biol Cell"},{"issue":"4","key":"pone.0330818.ref025","doi-asserted-by":"crossref","first-page":"299","DOI":"10.3109\/10799893.2011.587821","article-title":"Insulin targets the Na(+)\/K(+) ATPase in enterocytes via PI3K, PKC, and MAPKS","volume":"31","author":"MF Serhan","year":"2011","journal-title":"J Recept Signal Transduct Res"},{"key":"pone.0330818.ref026","doi-asserted-by":"crossref","DOI":"10.1152\/ajpcell.1995.268.5.C1277","article-title":"Protein kinase C-dependent stimulation of Na(+)-K(+)-ATP epsilon in rat proximal convoluted tubules","volume":"268","author":"E F\u00e9raille","year":"1995","journal-title":"Am J Physiol"},{"issue":"7","key":"pone.0330818.ref027","doi-asserted-by":"crossref","first-page":"1433","DOI":"10.1038\/sj.bjp.0703452","article-title":"Protein kinase C isoenzymes: a review of their structure, regulation and role in regulating airways smooth muscle tone and mitogenesis","volume":"130","author":"BL Webb","year":"2000","journal-title":"Br J Pharmacol"},{"issue":"3","key":"pone.0330818.ref028","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1016\/j.juro.2008.10.140","article-title":"S1P\/S1P2 signaling induces cyclooxygenase-2 expression in Wilms tumor","volume":"181","author":"M-H Li","year":"2009","journal-title":"J Urol"},{"issue":"9","key":"pone.0330818.ref029","doi-asserted-by":"crossref","first-page":"2506","DOI":"10.1172\/JCI31123","article-title":"Essential role of sphingosine 1-phosphate receptor 2 in pathological angiogenesis of the mouse retina","volume":"117","author":"A Skoura","year":"2007","journal-title":"J Clin Invest"},{"issue":"4","key":"pone.0330818.ref030","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1016\/0090-6980(90)90118-F","article-title":"Regulation of prostaglandin E2 synthesis in human amnion by protein kinase C","volume":"39","author":"J Sander","year":"1990","journal-title":"Prostaglandins"},{"issue":"11","key":"pone.0330818.ref031","doi-asserted-by":"crossref","first-page":"2005","DOI":"10.1681\/ASN.V9112005","article-title":"High glucose increases prostaglandin E2 synthesis in human peritoneal mesothelial cells","volume":"9","author":"T Sitter","year":"1998","journal-title":"Journal of the American Society of Nephrology"},{"issue":"1","key":"pone.0330818.ref032","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1007\/BF00675628","article-title":"Protein kinase C mediates flow-induced prostaglandin E2 production in osteoblasts","volume":"52","author":"KM Reich","year":"1993","journal-title":"Calcif Tissue Int"},{"issue":"5","key":"pone.0330818.ref033","first-page":"931","article-title":"Stimulation of prostaglandin production in rat thymic epithelial cells by protein kinase C mediated activation of phospholipase A2","volume":"27","author":"PS Liu","year":"1992","journal-title":"Biochem Int"},{"key":"pone.0330818.ref034","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/1471-2199-3-16","article-title":"Cyclooxygenase-2 is a neuronal target gene of NF-kappaB","volume":"3","author":"B Kaltschmidt","year":"2002","journal-title":"BMC Mol Biol"},{"issue":"12","key":"pone.0330818.ref035","doi-asserted-by":"crossref","first-page":"3375","DOI":"10.1111\/j.0953-816X.2004.03441.x","article-title":"Spinal NF-kB activation induces COX-2 upregulation and contributes to inflammatory pain hypersensitivity","volume":"19","author":"K-M Lee","year":"2004","journal-title":"Eur J Neurosci"},{"issue":"4","key":"pone.0330818.ref036","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1046\/j.1365-2567.1998.00646.x","article-title":"Specific NF\u2010\u03baB blockade selectively inhibits tumour necrosis factor\u2010\u03b1\u2010induced COX\u20102 but not constitutive COX\u20101 gene expression in HT\u201029 cells","volume":"95","author":"C Jobin","year":"1998","journal-title":"Immunology"},{"issue":"12","key":"pone.0330818.ref037","doi-asserted-by":"crossref","first-page":"1647","DOI":"10.1084\/jem.20020408","article-title":"Protein kinase C beta controls nuclear factor kappaB activation in B cells through selective regulation of the IkappaB kinase alpha","volume":"195","author":"K Saijo","year":"2002","journal-title":"J Exp Med"},{"issue":"10","key":"pone.0330818.ref038","doi-asserted-by":"crossref","first-page":"1767","DOI":"10.1006\/jmcc.2000.1211","article-title":"Angiotensin II induces nuclear factor- kappa B activation in cultured neonatal rat cardiomyocytes through protein kinase C signaling pathway","volume":"32","author":"P Rouet-Benzineb","year":"2000","journal-title":"J Mol Cell Cardiol"},{"issue":"4","key":"pone.0330818.ref039","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0095566","article-title":"Sphingosine 1-phosphate (S1P) induced interleukin-8 (IL-8) release is mediated by S1P receptor 2 and nuclear factor \u03baB in BEAS-2B cells","volume":"9","author":"MJ O\u2019Sullivan","year":"2014","journal-title":"PLoS One"},{"issue":"24","key":"pone.0330818.ref040","doi-asserted-by":"crossref","first-page":"22907","DOI":"10.1074\/jbc.M500130200","article-title":"Prostaglandin E2 stimulates fibronectin expression through EP1 receptor, phospholipase C, protein kinase Calpha, and c-Src pathway in primary cultured rat osteoblasts","volume":"280","author":"C-H Tang","year":"2005","journal-title":"J Biol 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Vo","year":"2013","journal-title":"Endocrinology"},{"issue":"7","key":"pone.0330818.ref044","article-title":"Gastrin decreases Na+,K+-ATPase activity via a PI 3-kinase- and PKC-dependent pathway in human renal proximal tubule cells","volume":"310","author":"T Liu","year":"2016","journal-title":"Am J Physiol Endocrinol Metab"},{"issue":"4","key":"pone.0330818.ref045","doi-asserted-by":"crossref","DOI":"10.1152\/ajpgi.00584.2006","article-title":"PGE2 inhibits apoptosis in human adenocarcinoma Caco-2 cell line through Ras-PI3K association and cAMP-dependent kinase A activation","volume":"293","author":"V Leone","year":"2007","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"issue":"18","key":"pone.0330818.ref046","doi-asserted-by":"crossref","first-page":"6958","DOI":"10.1021\/bi050119q","article-title":"The role of G beta gamma and domain interfaces in the activation of G protein-coupled receptor kinase 2","volume":"44","author":"DT 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