title: Calcium-mediated oxidative stress: a common mechanism in tight junction disruption by different types of cellular stress.
abstract: The role of reactive oxygen species (ROS) in osmotic stress, dextran sulfate sodium (DSS) and cyclic stretch-induced tight junction (TJ) disruption was investigated in Caco-2 cell monolayersin vitroand restraint stress-induced barrier dysfunction in mouse colonin vivoLive cell imaging showed that osmotic stress, cyclic stretch and DSS triggered rapid production of ROS in Caco-2 cell monolayers, which was blocked by depletion of intracellular Ca2+by 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. Knockdown of CaV1.3 or TRPV6 channels blocked osmotic stress and DSS-induced ROS production and attenuated TJ disruption and barrier dysfunction.N-Acetyl l-cysteine (NAC) and l-NG-Nitroarginine methyl ester (l-NAME) blocked stress-induced TJ disruption and barrier dysfunction. NAC and l-NAME also blocked stress-induced activation of c-JunN-terminal kinase (JNK) and c-Src. ROS was colocalized with the mitochondrial marker in stressed cells. Cyclosporin A blocked osmotic stress and DSS-induced ROS production, barrier dysfunction, TJ disruption and JNK activation. Mitochondria-targeted Mito-TEMPO blocked osmotic stress and DSS-induced barrier dysfunction and TJ disruption. Chronic restraint stress in mice resulted in the elevation of intracellular Ca2+, activation of JNK and c-Src, and disruption of TJ in the colonic epithelium. Furthermore, corticosterone administration induced JNK and c-Src activation, TJ disruption and protein thiol oxidation in colonic mucosa. The present study demonstrates that oxidative stress is a common signal in the mechanism of TJ disruption in the intestinal epithelium by different types of cellular stressin vitroand bio behavioral stressin vivo.
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, U.S.A.
authors.forename: Ruchika
authors.initials: R
authors.lastname: Gangwar
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, U.S.A.
authors.forename: Avtar S
authors.initials: AS
authors.lastname: Meena
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, U.S.A.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Shukla
authors.affiliation: Department of Gynecologic Oncology and Center for RNA Interference and Non-Coding RNA, M.D. Anderson Cancer Center, Houston, TX, U.S.A.
authors.forename: Archana S
authors.initials: AS
authors.lastname: Nagaraja
authors.affiliation: Department of Gynecologic Oncology and Center for RNA Interference and Non-Coding RNA, M.D. Anderson Cancer Center, Houston, TX, U.S.A.
authors.forename: Piotr L
authors.initials: PL
authors.lastname: Dorniak
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, U.S.A.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Pallikuth
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, U.S.A.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Waters
authors.affiliation: Department of Gynecologic Oncology and Center for RNA Interference and Non-Coding RNA, M.D. Anderson Cancer Center, Houston, TX, U.S.A.
authors.forename: Anil
authors.initials: A
authors.lastname: Sood
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, U.S.A. rrao2@uthsc.edu.
authors.forename: RadhaKrishna
authors.initials: R
authors.lastname: Rao
mesh_terms: D000818:Animals; D000081247:CSK Tyrosine-Protein Kinase; D018938:Caco-2 Cells; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000235:genetics / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003345:Corticosterone / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology; D016264:Dextran Sulfate / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D005786:Gene Expression Regulation; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D008297:Male; D040542:Mechanotransduction, Cellular; D051379:Mice; D008810:Mice, Inbred C57BL; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009997:Osmotic Pressure / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000819:agonists / Q000378:metabolism*; D013314:Stress, Mechanical; D013315:Stress, Psychological / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: C485995:Cacna1d protein, mouse; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D002614:Chelating Agents; D017382:Reactive Oxygen Species; D050916:TRPV Cation Channels; C446923:Trpv6 protein, mouse; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D016572:Cyclosporine; D016264:Dextran Sulfate; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human; D048031:JNK Mitogen-Activated Protein Kinases; D002118:Calcium; D019331:NG-Nitroarginine Methyl Ester; D003345:Corticosterone
keywords: ROS; adherens junction; barrier function; epithelium; intestine; tight junction