Rapid Effects of Dexamethasone on Intracellular pH and Na+/H+ Exchanger Activity in Human Bronchial Epithelial Cells
Glucocorticoids have been shown to produce rapid nongenomic responses in airway epithelia. By using an intracellular pH (pHi) spectrofluorescence imaging system and the NH4 Cl acid-loading technique, we have shown that the synthetic glucocorticoid,dexamethasone, accelerated intracellular pH recovery...
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| Veröffentlicht in: | The Journal of biological chemistry 2005-10, Vol.280 (43), p.35807-35814 |
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| creator | Verrière, Valia A. Hynes, Darina Faherty, Sheila Devaney, James Bousquet, Jean Harvey, Brian J. Urbach, Valérie |
| description | Glucocorticoids have been shown to produce rapid nongenomic responses in airway epithelia. By using an intracellular pH (pHi) spectrofluorescence imaging system and the NH4 Cl acid-loading technique, we have shown that the synthetic glucocorticoid,dexamethasone, accelerated intracellular pH recovery after an acid load in a human bronchial epithelial cell line (16HBE14o– cells). Exposure to NH4Cl (20 mm) elicited an intracellular acidification, followed by a pHi recovery. Inhibition of the Na+/H+ exchanger decreased the steady-state pHi and antagonized the dexamethasone stimulation of pHi regulation. The rapid effect of dexamethasone on pHi was neither affected by the inhibitor of transcription, cycloheximide, nor by the classical glucocorticoid and mineralocorticoid receptors antagonists, RU486 and spironolactone, respectively. The dexamethasone effect on pHi regulation was reduced by inhibitors of adenylate cyclase, cAMP-dependent protein kinase and mitogenactivated protein kinase (ERK1/2). By using a PepTag assay system and Western blotting, we have shown that dexamethasone stimulated cAMP-dependent protein kinase and mitogen-activated protein kinase activities. Taken together our results provide evidence for the rapid stimulation of Na+/H+ exchange activity by glucocorticoids in bronchial epithelial cells via a nongenomic mechanism involving cAMP-dependent protein kinase and mitogen-activated protein kinase ERK1/2 pathways. |
| doi_str_mv | 10.1074/jbc.M506584200 |
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By using an intracellular pH (pHi) spectrofluorescence imaging system and the NH4 Cl acid-loading technique, we have shown that the synthetic glucocorticoid,dexamethasone, accelerated intracellular pH recovery after an acid load in a human bronchial epithelial cell line (16HBE14o– cells). Exposure to NH4Cl (20 mm) elicited an intracellular acidification, followed by a pHi recovery. Inhibition of the Na+/H+ exchanger decreased the steady-state pHi and antagonized the dexamethasone stimulation of pHi regulation. The rapid effect of dexamethasone on pHi was neither affected by the inhibitor of transcription, cycloheximide, nor by the classical glucocorticoid and mineralocorticoid receptors antagonists, RU486 and spironolactone, respectively. The dexamethasone effect on pHi regulation was reduced by inhibitors of adenylate cyclase, cAMP-dependent protein kinase and mitogenactivated protein kinase (ERK1/2). By using a PepTag assay system and Western blotting, we have shown that dexamethasone stimulated cAMP-dependent protein kinase and mitogen-activated protein kinase activities. Taken together our results provide evidence for the rapid stimulation of Na+/H+ exchange activity by glucocorticoids in bronchial epithelial cells via a nongenomic mechanism involving cAMP-dependent protein kinase and mitogen-activated protein kinase ERK1/2 pathways.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M506584200</identifier><identifier>PMID: 16040608</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Anti-Inflammatory Agents - pharmacology ; Biochemistry ; Biochemistry, Molecular Biology ; Biophysics ; Bronchi - cytology ; Bronchi - pathology ; Calcium - metabolism ; Cell Line ; Cells, Cultured ; Cellular Biology ; Cyclic AMP - metabolism ; Cyclic AMP-Dependent Protein Kinases - metabolism ; Dexamethasone - pharmacology ; Epithelial Cells - cytology ; Glucocorticoids - metabolism ; Human health and pathology ; Humans ; Hydrogen-Ion Concentration ; Immunology ; Innate immunity ; Life Sciences ; MAP Kinase Signaling System ; Microbiology and Parasitology ; Mifepristone - pharmacology ; Mineralocorticoids - metabolism ; Mitogen-Activated Protein Kinase 1 - metabolism ; Mitogen-Activated Protein Kinase 3 - metabolism ; Pertussis Toxin - pharmacology ; Pharmaceutical sciences ; Pharmacology ; Phosphorylation ; Pulmonology and respiratory tract ; Sodium-Hydrogen Exchangers - metabolism ; Spectrometry, Fluorescence ; Steroids - metabolism ; Subcellular Processes ; Time Factors ; Tissues and Organs</subject><ispartof>The Journal of biological chemistry, 2005-10, Vol.280 (43), p.35807-35814</ispartof><rights>2005 © 2005 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c445t-93c7eebd784e838d394559a5e858ff6b20d236ea282ea0f40cfe66fc5c2e5c7f3</citedby><cites>FETCH-LOGICAL-c445t-93c7eebd784e838d394559a5e858ff6b20d236ea282ea0f40cfe66fc5c2e5c7f3</cites><orcidid>0000-0002-4061-4766</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16040608$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-03110579$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Verrière, Valia A.</creatorcontrib><creatorcontrib>Hynes, Darina</creatorcontrib><creatorcontrib>Faherty, Sheila</creatorcontrib><creatorcontrib>Devaney, James</creatorcontrib><creatorcontrib>Bousquet, Jean</creatorcontrib><creatorcontrib>Harvey, Brian J.</creatorcontrib><creatorcontrib>Urbach, Valérie</creatorcontrib><title>Rapid Effects of Dexamethasone on Intracellular pH and Na+/H+ Exchanger Activity in Human Bronchial Epithelial Cells</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Glucocorticoids have been shown to produce rapid nongenomic responses in airway epithelia. By using an intracellular pH (pHi) spectrofluorescence imaging system and the NH4 Cl acid-loading technique, we have shown that the synthetic glucocorticoid,dexamethasone, accelerated intracellular pH recovery after an acid load in a human bronchial epithelial cell line (16HBE14o– cells). Exposure to NH4Cl (20 mm) elicited an intracellular acidification, followed by a pHi recovery. Inhibition of the Na+/H+ exchanger decreased the steady-state pHi and antagonized the dexamethasone stimulation of pHi regulation. The rapid effect of dexamethasone on pHi was neither affected by the inhibitor of transcription, cycloheximide, nor by the classical glucocorticoid and mineralocorticoid receptors antagonists, RU486 and spironolactone, respectively. The dexamethasone effect on pHi regulation was reduced by inhibitors of adenylate cyclase, cAMP-dependent protein kinase and mitogenactivated protein kinase (ERK1/2). By using a PepTag assay system and Western blotting, we have shown that dexamethasone stimulated cAMP-dependent protein kinase and mitogen-activated protein kinase activities. Taken together our results provide evidence for the rapid stimulation of Na+/H+ exchange activity by glucocorticoids in bronchial epithelial cells via a nongenomic mechanism involving cAMP-dependent protein kinase and mitogen-activated protein kinase ERK1/2 pathways.</description><subject>Anti-Inflammatory Agents - pharmacology</subject><subject>Biochemistry</subject><subject>Biochemistry, Molecular Biology</subject><subject>Biophysics</subject><subject>Bronchi - cytology</subject><subject>Bronchi - pathology</subject><subject>Calcium - metabolism</subject><subject>Cell Line</subject><subject>Cells, Cultured</subject><subject>Cellular Biology</subject><subject>Cyclic AMP - metabolism</subject><subject>Cyclic AMP-Dependent Protein Kinases - metabolism</subject><subject>Dexamethasone - pharmacology</subject><subject>Epithelial Cells - cytology</subject><subject>Glucocorticoids - metabolism</subject><subject>Human health and pathology</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>Immunology</subject><subject>Innate immunity</subject><subject>Life Sciences</subject><subject>MAP Kinase Signaling System</subject><subject>Microbiology and Parasitology</subject><subject>Mifepristone - pharmacology</subject><subject>Mineralocorticoids - metabolism</subject><subject>Mitogen-Activated Protein Kinase 1 - metabolism</subject><subject>Mitogen-Activated Protein Kinase 3 - metabolism</subject><subject>Pertussis Toxin - pharmacology</subject><subject>Pharmaceutical sciences</subject><subject>Pharmacology</subject><subject>Phosphorylation</subject><subject>Pulmonology and respiratory tract</subject><subject>Sodium-Hydrogen Exchangers - metabolism</subject><subject>Spectrometry, Fluorescence</subject><subject>Steroids - metabolism</subject><subject>Subcellular Processes</subject><subject>Time Factors</subject><subject>Tissues and Organs</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kdGL1DAQxoMo3nr66qPkQQQ5upc0TZs-rutqD1YFUfAtpOnkmqNNapKud_-9XXbxnpyXGYbffMzMh9BrStaUVMX1XavXXzgpuShyQp6gFSWCZYzTX0_RipCcZnXOxQV6EeMdWaKo6XN0QUtSkJKIFUrf1WQ7vDMGdIrYG_wR7tUIqVfRO8De4RuXgtIwDPOgAp4arFyHv6qr6-YK7-51r9wtBLzRyR5sesDW4WYelcMfgne6t2rAu8mmHoZjuV104kv0zKghwqtzvkQ_P-1-bJts_-3zzXazz3RR8JTVTFcAbVeJAgQTHasLzmvFQXBhTNnmpMtZCSoXOShiCqINlKXRXOfAdWXYJXp_0u3VIKdgRxUepFdWNpu9PPYIo5Twqj7QhX13Yqfgf88QkxxtPF6tHPg5ylJUtKppuYDrE6iDjzGA-adMiTx6IhdP5KMny8Cbs_LcjtA94mcTFuDteU172_-xAWRrve5hlLkgsmCScUGqBRMnDJaXHSwEGbUFp6FbRnSSnbf_W-EvLySleg</recordid><startdate>20051028</startdate><enddate>20051028</enddate><creator>Verrière, Valia A.</creator><creator>Hynes, Darina</creator><creator>Faherty, Sheila</creator><creator>Devaney, James</creator><creator>Bousquet, Jean</creator><creator>Harvey, Brian J.</creator><creator>Urbach, Valérie</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope><orcidid>https://orcid.org/0000-0002-4061-4766</orcidid></search><sort><creationdate>20051028</creationdate><title>Rapid Effects of Dexamethasone on Intracellular pH and Na+/H+ Exchanger Activity in Human Bronchial Epithelial Cells</title><author>Verrière, Valia A. ; Hynes, Darina ; Faherty, Sheila ; Devaney, James ; Bousquet, Jean ; Harvey, Brian J. ; Urbach, Valérie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c445t-93c7eebd784e838d394559a5e858ff6b20d236ea282ea0f40cfe66fc5c2e5c7f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Anti-Inflammatory Agents - pharmacology</topic><topic>Biochemistry</topic><topic>Biochemistry, Molecular Biology</topic><topic>Biophysics</topic><topic>Bronchi - cytology</topic><topic>Bronchi - pathology</topic><topic>Calcium - metabolism</topic><topic>Cell Line</topic><topic>Cells, Cultured</topic><topic>Cellular Biology</topic><topic>Cyclic AMP - metabolism</topic><topic>Cyclic AMP-Dependent Protein Kinases - metabolism</topic><topic>Dexamethasone - pharmacology</topic><topic>Epithelial Cells - cytology</topic><topic>Glucocorticoids - metabolism</topic><topic>Human health and pathology</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>Immunology</topic><topic>Innate immunity</topic><topic>Life Sciences</topic><topic>MAP Kinase Signaling System</topic><topic>Microbiology and Parasitology</topic><topic>Mifepristone - pharmacology</topic><topic>Mineralocorticoids - metabolism</topic><topic>Mitogen-Activated Protein Kinase 1 - metabolism</topic><topic>Mitogen-Activated Protein Kinase 3 - metabolism</topic><topic>Pertussis Toxin - pharmacology</topic><topic>Pharmaceutical sciences</topic><topic>Pharmacology</topic><topic>Phosphorylation</topic><topic>Pulmonology and respiratory tract</topic><topic>Sodium-Hydrogen Exchangers - metabolism</topic><topic>Spectrometry, Fluorescence</topic><topic>Steroids - metabolism</topic><topic>Subcellular Processes</topic><topic>Time Factors</topic><topic>Tissues and Organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Verrière, Valia A.</creatorcontrib><creatorcontrib>Hynes, Darina</creatorcontrib><creatorcontrib>Faherty, Sheila</creatorcontrib><creatorcontrib>Devaney, James</creatorcontrib><creatorcontrib>Bousquet, Jean</creatorcontrib><creatorcontrib>Harvey, Brian J.</creatorcontrib><creatorcontrib>Urbach, Valérie</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Verrière, Valia A.</au><au>Hynes, Darina</au><au>Faherty, Sheila</au><au>Devaney, James</au><au>Bousquet, Jean</au><au>Harvey, Brian J.</au><au>Urbach, Valérie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid Effects of Dexamethasone on Intracellular pH and Na+/H+ Exchanger Activity in Human Bronchial Epithelial Cells</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2005-10-28</date><risdate>2005</risdate><volume>280</volume><issue>43</issue><spage>35807</spage><epage>35814</epage><pages>35807-35814</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Glucocorticoids have been shown to produce rapid nongenomic responses in airway epithelia. By using an intracellular pH (pHi) spectrofluorescence imaging system and the NH4 Cl acid-loading technique, we have shown that the synthetic glucocorticoid,dexamethasone, accelerated intracellular pH recovery after an acid load in a human bronchial epithelial cell line (16HBE14o– cells). Exposure to NH4Cl (20 mm) elicited an intracellular acidification, followed by a pHi recovery. Inhibition of the Na+/H+ exchanger decreased the steady-state pHi and antagonized the dexamethasone stimulation of pHi regulation. The rapid effect of dexamethasone on pHi was neither affected by the inhibitor of transcription, cycloheximide, nor by the classical glucocorticoid and mineralocorticoid receptors antagonists, RU486 and spironolactone, respectively. The dexamethasone effect on pHi regulation was reduced by inhibitors of adenylate cyclase, cAMP-dependent protein kinase and mitogenactivated protein kinase (ERK1/2). By using a PepTag assay system and Western blotting, we have shown that dexamethasone stimulated cAMP-dependent protein kinase and mitogen-activated protein kinase activities. Taken together our results provide evidence for the rapid stimulation of Na+/H+ exchange activity by glucocorticoids in bronchial epithelial cells via a nongenomic mechanism involving cAMP-dependent protein kinase and mitogen-activated protein kinase ERK1/2 pathways.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>16040608</pmid><doi>10.1074/jbc.M506584200</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0002-4061-4766</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Anti-Inflammatory Agents - pharmacology Biochemistry Biochemistry, Molecular Biology Biophysics Bronchi - cytology Bronchi - pathology Calcium - metabolism Cell Line Cells, Cultured Cellular Biology Cyclic AMP - metabolism Cyclic AMP-Dependent Protein Kinases - metabolism Dexamethasone - pharmacology Epithelial Cells - cytology Glucocorticoids - metabolism Human health and pathology Humans Hydrogen-Ion Concentration Immunology Innate immunity Life Sciences MAP Kinase Signaling System Microbiology and Parasitology Mifepristone - pharmacology Mineralocorticoids - metabolism Mitogen-Activated Protein Kinase 1 - metabolism Mitogen-Activated Protein Kinase 3 - metabolism Pertussis Toxin - pharmacology Pharmaceutical sciences Pharmacology Phosphorylation Pulmonology and respiratory tract Sodium-Hydrogen Exchangers - metabolism Spectrometry, Fluorescence Steroids - metabolism Subcellular Processes Time Factors Tissues and Organs |
| title | Rapid Effects of Dexamethasone on Intracellular pH and Na+/H+ Exchanger Activity in Human Bronchial Epithelial Cells |
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