title: Flufenamic acid protects against intestinal fluid secretion and barrier leakage in a mouse model of Vibrio cholerae infection through NF-κB inhibition and AMPK activation.
abstract: Nuclear factor kappa B (NF-κB)-mediated inflammatory responses play crucial roles in the pathogenesis of diarrhea caused by the Vibrio cholerae El Tor variant (EL), which is a major bacterial strain causing recent cholera outbreaks. Flufenamic acid (FFA) has previously been demonstrated to be a potent activator of AMP-activated protein kinase (AMPK), which is a negative regulator of NF-κB signaling. This study aimed to investigate the anti-diarrheal efficacy of FFA in a mouse model of EL infection and to investigate the mechanisms by which FFA activates AMPK in intestinal epithelial cells (IEC). In a mouse closed loop model of EL infection, FFA treatment (20mg/kg) significantly abrogated EL-induced intestinal fluid secretion and barrier disruption. In addition, FFA suppressed NF-κB nuclear translocation and expression of proinflammatory mediators and promoted AMPK phosphorylation in the EL-infected mouse intestine. In T84 cells, FFA induced AMPK activation. Furthermore, FFA promoted tight junction assembly and prevented interferon gamma (IFN-γ)-induced barrier disruption in an AMPK-dependent manner. Biochemical and molecular docking analyses indicated that FFA activates AMPK via a direct stimulation of calcium/calmodulin-dependent protein kinase kinase beta (CaMKKβ) activity. Collectively, our data indicate that FFA represents a class of existing drugs that may be of potential utility in the treatment of cholera caused by EL infection via AMPK-mediated suppression of NF-κB signaling in IEC.
authors.affiliation: Graduate Program in Translational Medicine, Research Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand.
authors.forename: Pawin
authors.initials: P
authors.lastname: Pongkorpsakol
authors.affiliation: Department of Physiology, Faculty of Science, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand.
authors.forename: Saravut
authors.initials: S
authors.lastname: Satitsri
authors.affiliation: Department of Physiology, Faculty of Science, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand.
authors.forename: Preedajit
authors.initials: P
authors.lastname: Wongkrasant
authors.affiliation: Department of Pharmacology, Faculty of Science, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand.
authors.forename: Pamorn
authors.initials: P
authors.lastname: Chittavanich
authors.affiliation: Department of Basic Medical Science, Faculty of Medicine Vajira Hospital, Navamindradhiraj University, Thailand.
authors.forename: Suticha
authors.initials: S
authors.lastname: Kittayaruksakul
authors.affiliation: Graduate Studies, Faculty of Allied Health Science, Thammasat University, Paholyothin Road, Rangsit, Prathumthani 12120, Thailand.
authors.forename: Potjanee
authors.initials: P
authors.lastname: Srimanote
authors.affiliation: Department of Physiology, Faculty of Science, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand.
authors.forename: Varanuj
authors.initials: V
authors.lastname: Chatsudthipong
authors.affiliation: Department of Physiology, Faculty of Science, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand; Excellent Center for Drug Discovery, Mahidol University, Rama VI Road, Rajathevi, Bangkok 10400, Thailand. Electronic address: chatchai.mua@mahidol.ac.th.
authors.forename: Chatchai
authors.initials: C
authors.lastname: Muanprasat
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000818:Animals; D001826:Body Fluids / Q000187:drug effects / Q000378:metabolism*; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000737:chemistry / Q000378:metabolism; D020134:Catalytic Domain; D002460:Cell Line; D002771:Cholera / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism; D003967:Diarrhea / Q000188:drug therapy / Q000821:virology; D004195:Disease Models, Animal; D005439:Flufenamic Acid / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007422:Intestines / Q000187:drug effects*; D051379:Mice; D062105:Molecular Docking Simulation; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D014734:Vibrio cholerae / Q000502:physiology*
chemical_list: D016328:NF-kappa B; D005439:Flufenamic Acid; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D055372:AMP-Activated Protein Kinases
keywords: AMP-activated protein kinase; Cholera; Flufenamic acid; Inflammation; Vibrio cholerae