Thymoquinone suppressses in vitro production of IL-5 and IL-13 by mast cells in response to lipopolysaccharide stimulation
Activated mast cells produce Th2 cytokines that regulate allergic inflammation. We have previously shown that thymoquinone (TQ) attenuated airway inflammation in a mouse model of allergic airway inflammation. The present study investigated whether TQ affects Th2 cytokine response in vitro in lipopol...
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| Veröffentlicht in: | Inflammation research 2007-08, Vol.56 (8), p.345-351 |
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| description | Activated mast cells produce Th2 cytokines that regulate allergic inflammation. We have previously shown that thymoquinone (TQ) attenuated airway inflammation in a mouse model of allergic airway inflammation. The present study investigated whether TQ affects Th2 cytokine response in vitro in lipopolysaccharide (LPS)-activated rat mast cells, RBL-2H3.
RBL-2H3 cells were stimulated for 12 h with 0.1 microg/ml LPS in the presence or absence of 10 microM TQ. Th2 cytokine production was measured in the culture supernatants by ELISA. The mRNA expression of IL-5, IL- 13 and GATA transcription factors was determined by RT-PCR. The expression of the transcription proteins c-Fos, c- Jun and phospho-c-Jun were determined by western blotting. The in vivo binding of GATA, AP-1 and NF-AT transcription factors to IL-5 promoter was assessed by chromatin immunoprecipitation analysis.
TQ significantly (p |
| doi_str_mv | 10.1007/s00011-007-7051-0 |
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RBL-2H3 cells were stimulated for 12 h with 0.1 microg/ml LPS in the presence or absence of 10 microM TQ. Th2 cytokine production was measured in the culture supernatants by ELISA. The mRNA expression of IL-5, IL- 13 and GATA transcription factors was determined by RT-PCR. The expression of the transcription proteins c-Fos, c- Jun and phospho-c-Jun were determined by western blotting. The in vivo binding of GATA, AP-1 and NF-AT transcription factors to IL-5 promoter was assessed by chromatin immunoprecipitation analysis.
TQ significantly (p <0.05) inhibited LPS-induced IL-5 and IL-13 mRNA expression and protein production. However, TQ did not affect IL-10 production. GATA transcription factors are involved in the transcription of IL-5 and IL-13. TQ had no effect on the expression of AP-1 protein subunits, c-Jun and c-Fos, but markedly reduced the transcription of GATA-1 and -2 genes. Chromatin immunoprecipitation revealed that GATA, AP-1 and NF-AT binding to IL-5 promoter was induced by LPS stimulation and that TQ inhibited GATA binding at the IL-5 promoter but did not affect AP-1 and NF-AT binding.
These results suggest that TQ inhibits LPS-induced proinflammatory cytokine production in RBL-2H3 cells by blocking GATA transcription factor expression and promoter binding which demonstrates the anti-inflammatory effect of TQ.</description><identifier>ISSN: 1023-3830</identifier><identifier>EISSN: 1420-908X</identifier><identifier>DOI: 10.1007/s00011-007-7051-0</identifier><identifier>PMID: 17687519</identifier><language>eng</language><publisher>Switzerland: Springer Nature B.V</publisher><subject>Animals ; Anti-Inflammatory Agents - pharmacology ; Benzoquinones - pharmacology ; Cells, Cultured ; GATA1 Transcription Factor - genetics ; GATA2 Transcription Factor - genetics ; Interleukin-13 - biosynthesis ; Interleukin-5 - biosynthesis ; Interleukin-5 - genetics ; Lipopolysaccharides - pharmacology ; Mast Cells - drug effects ; Mast Cells - metabolism ; NF-kappa B - metabolism ; Promoter Regions, Genetic ; Rats ; Transcription Factor AP-1 - metabolism</subject><ispartof>Inflammation research, 2007-08, Vol.56 (8), p.345-351</ispartof><rights>Birkhäuser Verlag, Basel 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c357t-ab03b441da4dc86cca7d3ecc634d7c063062da2d43b182fe2649967d1ae4ed6d3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17687519$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>El Gazzar, M A</creatorcontrib><title>Thymoquinone suppressses in vitro production of IL-5 and IL-13 by mast cells in response to lipopolysaccharide stimulation</title><title>Inflammation research</title><addtitle>Inflamm Res</addtitle><description>Activated mast cells produce Th2 cytokines that regulate allergic inflammation. We have previously shown that thymoquinone (TQ) attenuated airway inflammation in a mouse model of allergic airway inflammation. The present study investigated whether TQ affects Th2 cytokine response in vitro in lipopolysaccharide (LPS)-activated rat mast cells, RBL-2H3.
RBL-2H3 cells were stimulated for 12 h with 0.1 microg/ml LPS in the presence or absence of 10 microM TQ. Th2 cytokine production was measured in the culture supernatants by ELISA. The mRNA expression of IL-5, IL- 13 and GATA transcription factors was determined by RT-PCR. The expression of the transcription proteins c-Fos, c- Jun and phospho-c-Jun were determined by western blotting. The in vivo binding of GATA, AP-1 and NF-AT transcription factors to IL-5 promoter was assessed by chromatin immunoprecipitation analysis.
TQ significantly (p <0.05) inhibited LPS-induced IL-5 and IL-13 mRNA expression and protein production. However, TQ did not affect IL-10 production. GATA transcription factors are involved in the transcription of IL-5 and IL-13. TQ had no effect on the expression of AP-1 protein subunits, c-Jun and c-Fos, but markedly reduced the transcription of GATA-1 and -2 genes. Chromatin immunoprecipitation revealed that GATA, AP-1 and NF-AT binding to IL-5 promoter was induced by LPS stimulation and that TQ inhibited GATA binding at the IL-5 promoter but did not affect AP-1 and NF-AT binding.
These results suggest that TQ inhibits LPS-induced proinflammatory cytokine production in RBL-2H3 cells by blocking GATA transcription factor expression and promoter binding which demonstrates the anti-inflammatory effect of TQ.</description><subject>Animals</subject><subject>Anti-Inflammatory Agents - pharmacology</subject><subject>Benzoquinones - pharmacology</subject><subject>Cells, Cultured</subject><subject>GATA1 Transcription Factor - genetics</subject><subject>GATA2 Transcription Factor - genetics</subject><subject>Interleukin-13 - biosynthesis</subject><subject>Interleukin-5 - biosynthesis</subject><subject>Interleukin-5 - genetics</subject><subject>Lipopolysaccharides - pharmacology</subject><subject>Mast Cells - drug effects</subject><subject>Mast Cells - metabolism</subject><subject>NF-kappa B - metabolism</subject><subject>Promoter Regions, Genetic</subject><subject>Rats</subject><subject>Transcription Factor AP-1 - metabolism</subject><issn>1023-3830</issn><issn>1420-908X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNqFkUtr3TAQhUVJaNK0P6CbIrLozs3oYUleltBH4EI3CWQnZEmXKNiWo7ELt7--cu-FQjddzVl858wMh5D3DD4xAH2DAMBYU2Wjoa3iFblkkkPTgXk8qxq4aIQRcEHeID5X2nDDX5MLppXRLesuya_7p8OYX9Y05SlSXOe5RESMSNNEf6alZDqXHFa_pDzRvKd3u6albgqbYIL2Bzo6XKiPw_DHU-1znjDSJdMhzXnOwwGd90-upFA3LGlcB7elvSXnezdgfHeaV-Th65f72-_N7se3u9vPu8aLVi-N60H0UrLgZPBGee90ENF7JWTQHpQAxYPjQYqeGb6PXMmuUzowF2UMKogr8vGYWx95WSMudky43eummFe0yjBZDfq_IAfZgVSigtf_gM95LVN9wnKmuDBMtRViR8iXjFji3s4lja4cLAO71WeP9dlNbvVZqJ4Pp-C1H2P46zj1JX4DMfWW0w</recordid><startdate>200708</startdate><enddate>200708</enddate><creator>El Gazzar, M A</creator><general>Springer Nature B.V</general><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>3V.</scope><scope>7QL</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>200708</creationdate><title>Thymoquinone suppressses in vitro production of IL-5 and IL-13 by mast cells in response to lipopolysaccharide stimulation</title><author>El Gazzar, M A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c357t-ab03b441da4dc86cca7d3ecc634d7c063062da2d43b182fe2649967d1ae4ed6d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Anti-Inflammatory Agents - pharmacology</topic><topic>Benzoquinones - pharmacology</topic><topic>Cells, Cultured</topic><topic>GATA1 Transcription Factor - genetics</topic><topic>GATA2 Transcription Factor - genetics</topic><topic>Interleukin-13 - biosynthesis</topic><topic>Interleukin-5 - biosynthesis</topic><topic>Interleukin-5 - genetics</topic><topic>Lipopolysaccharides - pharmacology</topic><topic>Mast Cells - drug effects</topic><topic>Mast Cells - metabolism</topic><topic>NF-kappa B - metabolism</topic><topic>Promoter Regions, Genetic</topic><topic>Rats</topic><topic>Transcription Factor AP-1 - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>El Gazzar, M A</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>Inflammation research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>El Gazzar, M A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Thymoquinone suppressses in vitro production of IL-5 and IL-13 by mast cells in response to lipopolysaccharide stimulation</atitle><jtitle>Inflammation research</jtitle><addtitle>Inflamm Res</addtitle><date>2007-08</date><risdate>2007</risdate><volume>56</volume><issue>8</issue><spage>345</spage><epage>351</epage><pages>345-351</pages><issn>1023-3830</issn><eissn>1420-908X</eissn><abstract>Activated mast cells produce Th2 cytokines that regulate allergic inflammation. We have previously shown that thymoquinone (TQ) attenuated airway inflammation in a mouse model of allergic airway inflammation. The present study investigated whether TQ affects Th2 cytokine response in vitro in lipopolysaccharide (LPS)-activated rat mast cells, RBL-2H3.
RBL-2H3 cells were stimulated for 12 h with 0.1 microg/ml LPS in the presence or absence of 10 microM TQ. Th2 cytokine production was measured in the culture supernatants by ELISA. The mRNA expression of IL-5, IL- 13 and GATA transcription factors was determined by RT-PCR. The expression of the transcription proteins c-Fos, c- Jun and phospho-c-Jun were determined by western blotting. The in vivo binding of GATA, AP-1 and NF-AT transcription factors to IL-5 promoter was assessed by chromatin immunoprecipitation analysis.
TQ significantly (p <0.05) inhibited LPS-induced IL-5 and IL-13 mRNA expression and protein production. However, TQ did not affect IL-10 production. GATA transcription factors are involved in the transcription of IL-5 and IL-13. TQ had no effect on the expression of AP-1 protein subunits, c-Jun and c-Fos, but markedly reduced the transcription of GATA-1 and -2 genes. Chromatin immunoprecipitation revealed that GATA, AP-1 and NF-AT binding to IL-5 promoter was induced by LPS stimulation and that TQ inhibited GATA binding at the IL-5 promoter but did not affect AP-1 and NF-AT binding.
These results suggest that TQ inhibits LPS-induced proinflammatory cytokine production in RBL-2H3 cells by blocking GATA transcription factor expression and promoter binding which demonstrates the anti-inflammatory effect of TQ.</abstract><cop>Switzerland</cop><pub>Springer Nature B.V</pub><pmid>17687519</pmid><doi>10.1007/s00011-007-7051-0</doi><tpages>7</tpages></addata></record> |
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| subjects | Animals Anti-Inflammatory Agents - pharmacology Benzoquinones - pharmacology Cells, Cultured GATA1 Transcription Factor - genetics GATA2 Transcription Factor - genetics Interleukin-13 - biosynthesis Interleukin-5 - biosynthesis Interleukin-5 - genetics Lipopolysaccharides - pharmacology Mast Cells - drug effects Mast Cells - metabolism NF-kappa B - metabolism Promoter Regions, Genetic Rats Transcription Factor AP-1 - metabolism |
| title | Thymoquinone suppressses in vitro production of IL-5 and IL-13 by mast cells in response to lipopolysaccharide stimulation |
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