Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2
Identifieur interne : 000123 ( Pmc/Curation ); précédent : 000122; suivant : 000124Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2
Auteurs : Orsolya Farkas [Hongrie] ; Orsolya Pal Cz [Hongrie] ; Erzsébet Pászti-Gere [Hongrie] ; Péter Gálfi [Hongrie]Source :
- Oxidative Medicine and Cellular Longevity [ 1942-0900 ] ; 2015.
Abstract
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DOI: 10.1155/2015/673847
PubMed: 26180592
PubMed Central: 4477253
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2</title><author><name sortKey="Farkas, Orsolya" sort="Farkas, Orsolya" uniqKey="Farkas O" first="Orsolya" last="Farkas">Orsolya Farkas</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author><author><name sortKey="Pal Cz, Orsolya" sort="Pal Cz, Orsolya" uniqKey="Pal Cz O" first="Orsolya" last="Pal Cz">Orsolya Pal Cz</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author><author><name sortKey="Paszti Gere, Erzsebet" sort="Paszti Gere, Erzsebet" uniqKey="Paszti Gere E" first="Erzsébet" last="Pászti-Gere">Erzsébet Pászti-Gere</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author><author><name sortKey="Galfi, Peter" sort="Galfi, Peter" uniqKey="Galfi P" first="Péter" last="Gálfi">Péter Gálfi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26180592</idno><idno type="pmc">4477253</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4477253</idno><idno type="RBID">PMC:4477253</idno><idno type="doi">10.1155/2015/673847</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000123</idno><idno type="wicri:Area/Pmc/Curation">000123</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2</title><author><name sortKey="Farkas, Orsolya" sort="Farkas, Orsolya" uniqKey="Farkas O" first="Orsolya" last="Farkas">Orsolya Farkas</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author><author><name sortKey="Pal Cz, Orsolya" sort="Pal Cz, Orsolya" uniqKey="Pal Cz O" first="Orsolya" last="Pal Cz">Orsolya Pal Cz</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author><author><name sortKey="Paszti Gere, Erzsebet" sort="Paszti Gere, Erzsebet" uniqKey="Paszti Gere E" first="Erzsébet" last="Pászti-Gere">Erzsébet Pászti-Gere</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author><author><name sortKey="Galfi, Peter" sort="Galfi, Peter" uniqKey="Galfi P" first="Péter" last="Gálfi">Péter Gálfi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</nlm:aff><country xml:lang="fr">Hongrie</country><wicri:regionArea>Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078</wicri:regionArea></affiliation></author></analytic><series><title level="j">Oxidative Medicine and Cellular Longevity</title><idno type="ISSN">1942-0900</idno><idno type="eISSN">1942-0994</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The <italic>in vitro</italic> anti-inflammatory effect of apigenin and its trimethylated analogue (apigenin-trimethylether) has been investigated in order to evaluate whether these flavonoids could attenuate LPS-induced inflammation in IPEC-J2 non-transformed intestinal epithelial cells. Levels of IL-6, IL-8, TNF-<italic>α</italic>, and COX-2 mRNA were measured as a marker of inflammatory response. The extracellular H<sub>2</sub>O<sub>2</sub> level in IPEC-J2 cells was also monitored by Amplex Red assay. Our data revealed that both compounds had significant lowering effect on the inflammatory response. Apigenin (at 25 <italic>μ</italic>M) significantly decreased gene expression of IL-6 in LPS-treated cells, while apigenin-trimethylether in the same concentration did not influence IL-6 mRNA level. Both apigenin and apigenin-trimethylether reduced IL-8 gene expression significantly. TNF-<italic>α</italic> mRNA level was decreased by apigenin-trimethylether, which was not influenced by apigenin. Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one. Furthermore, both flavonoids reduced significantly the level of extracellular H<sub>2</sub>O<sub>2</sub> compared to the control cells. In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Bisht, K" uniqKey="Bisht K">K. Bisht</name></author><author><name sortKey="Wagner, K H" uniqKey="Wagner K">K.-H. Wagner</name></author><author><name sortKey="Bulmer, A C" uniqKey="Bulmer A">A. C. Bulmer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Williams, R J" uniqKey="Williams R">R. J. Williams</name></author><author><name sortKey="Spencer, J P E" uniqKey="Spencer J">J. P. E. Spencer</name></author><author><name sortKey="Rice Evans, C" uniqKey="Rice Evans C">C. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Oxid Med Cell Longev</journal-id><journal-id journal-id-type="iso-abbrev">Oxid Med Cell Longev</journal-id><journal-id journal-id-type="publisher-id">OMCL</journal-id><journal-title-group><journal-title>Oxidative Medicine and Cellular Longevity</journal-title></journal-title-group><issn pub-type="ppub">1942-0900</issn><issn pub-type="epub">1942-0994</issn><publisher><publisher-name>Hindawi Publishing Corporation</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26180592</article-id><article-id pub-id-type="pmc">4477253</article-id><article-id pub-id-type="doi">10.1155/2015/673847</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Farkas</surname><given-names>Orsolya</given-names></name><xref ref-type="aff" rid="I1"></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author"><name><surname>Palócz</surname><given-names>Orsolya</given-names></name><xref ref-type="aff" rid="I1"></xref></contrib><contrib contrib-type="author"><name><surname>Pászti-Gere</surname><given-names>Erzsébet</given-names></name><xref ref-type="aff" rid="I1"></xref></contrib><contrib contrib-type="author"><name><surname>Gálfi</surname><given-names>Péter</given-names></name><xref ref-type="aff" rid="I1"></xref></contrib></contrib-group><aff id="I1">Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary</aff><author-notes><corresp id="cor1">*Orsolya Farkas: <email>farkas.orsolya@aotk.szie.hu</email></corresp><fn fn-type="other"><p>Academic Editor: Tullia Maraldi</p></fn></author-notes><pub-date pub-type="ppub"><year>2015</year></pub-date><pub-date pub-type="epub"><day>9</day><month>6</month><year>2015</year></pub-date><volume>2015</volume><elocation-id>673847</elocation-id><history><date date-type="received"><day>18</day><month>9</month><year>2014</year></date><date date-type="accepted"><day>10</day><month>12</month><year>2014</year></date></history><permissions><copyright-statement>Copyright © 2015 Orsolya Farkas et al.</copyright-statement><copyright-year>2015</copyright-year><license license-type="open-access"><license-p>This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><p>The <italic>in vitro</italic> anti-inflammatory effect of apigenin and its trimethylated analogue (apigenin-trimethylether) has been investigated in order to evaluate whether these flavonoids could attenuate LPS-induced inflammation in IPEC-J2 non-transformed intestinal epithelial cells. Levels of IL-6, IL-8, TNF-<italic>α</italic>, and COX-2 mRNA were measured as a marker of inflammatory response. The extracellular H<sub>2</sub>O<sub>2</sub> level in IPEC-J2 cells was also monitored by Amplex Red assay. Our data revealed that both compounds had significant lowering effect on the inflammatory response. Apigenin (at 25 <italic>μ</italic>M) significantly decreased gene expression of IL-6 in LPS-treated cells, while apigenin-trimethylether in the same concentration did not influence IL-6 mRNA level. Both apigenin and apigenin-trimethylether reduced IL-8 gene expression significantly. TNF-<italic>α</italic> mRNA level was decreased by apigenin-trimethylether, which was not influenced by apigenin. Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one. Furthermore, both flavonoids reduced significantly the level of extracellular H<sub>2</sub>O<sub>2</sub> compared to the control cells. In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.</p></abstract></article-meta></front><floats-group><fig id="fig1" orientation="portrait" position="float"><label>Figure 1</label><caption><p>Structure of apigenin (3′,4′,5-trihydroxyflavone) (a) and apigenin-trimethylether (3′,4′,5-trimethoxyflavone) (b).</p></caption><graphic xlink:href="OMCL2015-673847.001"></graphic></fig><fig id="fig2" orientation="portrait" position="float"><label>Figure 2</label><caption><p>Rate of living IPEC-J2 cells after apigenin and apigenin-trimethylether treatment. Cell viability was tested using Neutral Red uptake method. Effect of flavones on cell viability was studied in a dose- (a) and time-dependent (b) manner. Data are shown as means + standard deviations.</p></caption><graphic xlink:href="OMCL2015-673847.002"></graphic></fig><fig id="fig3" orientation="portrait" position="float"><label>Figure 3</label><caption><p>Relative gene expression of IL-6 in IPEC-J2 cells exposed to LPS treatment (at 1 <italic>μ</italic>g/mL; 1 h treatment). Effect of apigenin (25 <italic>μ</italic>M) and apigenin-trimethylether (25 <italic>μ</italic>M) on the IL-6 mRNA levels (<italic>n</italic> = 3-4/group; <sup>*</sup><italic>P</italic> < 0.05). Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.</p></caption><graphic xlink:href="OMCL2015-673847.003"></graphic></fig><fig id="fig4" orientation="portrait" position="float"><label>Figure 4</label><caption><p>Relative gene expression of IL-8 in IPEC-J2 cells exposed to LPS treatment (at 1 <italic>μ</italic>g/mL; 1 h treatment). Effect of apigenin (25 <italic>μ</italic>M) and apigenin-trimethylether (25 <italic>μ</italic>M) on the IL-8 mRNA levels (<italic>n</italic> = 3-4/group; <sup>*</sup><italic>P</italic> < 0.05). Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.</p></caption><graphic xlink:href="OMCL2015-673847.004"></graphic></fig><fig id="fig5" orientation="portrait" position="float"><label>Figure 5</label><caption><p>Relative gene expression of TNF-<italic>α</italic> in IPEC-J2 cells exposed to LPS treatment (at 1 <italic>μ</italic>g/mL; 1 h treatment). Effect of apigenin (25 <italic>μ</italic>M) and apigenin-trimethylether (25 <italic>μ</italic>M) on the TNF-<italic>α</italic> mRNA levels (<italic>n</italic> = 3-4/group; <sup>*</sup><italic>P</italic> < 0.05). Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.</p></caption><graphic xlink:href="OMCL2015-673847.005"></graphic></fig><fig id="fig6" orientation="portrait" position="float"><label>Figure 6</label><caption><p>Relative gene expression of COX-2 in IPEC-J2 cells exposed to LPS treatment (at 1 <italic>μ</italic>g/mL; 1 h treatment). Effect of apigenin (25 <italic>μ</italic>M) and apigenin-trimethylether (25 <italic>μ</italic>M) on the COX-2 mRNA levels (<italic>n</italic> = 3-4/group; <sup>*</sup><italic>P</italic> < 0.05). Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.</p></caption><graphic xlink:href="OMCL2015-673847.006"></graphic></fig><fig id="fig7" orientation="portrait" position="float"><label>Figure 7</label><caption><p>Level of extracellular H<sub>2</sub>O<sub>2</sub> in IPEC-J2 cells exposed to flavonoid treatment (25 and 50 <italic>μ</italic>M; 1 h). Effect of apigenin and apigenin-trimethylether on the relative extracellular H<sub>2</sub>O<sub>2</sub> levels (<italic>n</italic> = 3-4/group; <sup>*</sup><italic>P</italic> < 0.05). Fluorescence measurement was performed by Amplex Red method. Fluorescence was detected immediately (a) and 24 h after (b) flavonoid treatment. Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.</p></caption><graphic xlink:href="OMCL2015-673847.007"></graphic></fig><table-wrap id="tab1" orientation="portrait" position="float"><label>Table 1</label><caption><p>Sequence of primer sets used for quantitative real-time.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Gene symbol</th><th align="left" rowspan="1" colspan="1">Accession number</th><th align="left" rowspan="1" colspan="1">Primer sequences</th><th align="center" rowspan="1" colspan="1">Product size (bp)</th><th align="center" rowspan="1" colspan="1">Efficiency</th><th align="center" rowspan="1" colspan="1"><italic>R</italic><sup>2</sup></th><th align="center" rowspan="1" colspan="1">Reference</th></tr></thead><tbody><tr><td align="left" rowspan="2" colspan="1">IL-8</td><td align="left" rowspan="2" colspan="1">NM_213867</td><td align="left" rowspan="1" colspan="1">F 5′-AGAGGTCTGCCTGGACCCCA-3′</td><td align="center" rowspan="2" colspan="1">126</td><td align="center" rowspan="2" colspan="1">1.972</td><td align="center" rowspan="2" colspan="1">0.999</td><td align="center" rowspan="2" colspan="1">[<xref rid="B51" ref-type="bibr">28</xref>]</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-GGGAGCCACGGAGAATGGGT-3′</td></tr><tr><td align="center" colspan="7" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="2" colspan="1">IL-6</td><td align="left" rowspan="2" colspan="1">NM_214399</td><td align="left" rowspan="1" colspan="1">F 5′-TTCACCTCTCCGGACAAAAC-3′</td><td align="center" rowspan="2" colspan="1">122</td><td align="center" rowspan="2" colspan="1">1.970</td><td align="center" rowspan="2" colspan="1">0.995</td><td align="center" rowspan="2" colspan="1">[<xref rid="B52" ref-type="bibr">29</xref>]</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-TCTGCCAGTACCTCCTTGCT-3′</td></tr><tr><td align="center" colspan="7" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="2" colspan="1">TNF-<italic>α</italic></td><td align="left" rowspan="2" colspan="1">NM_214022</td><td align="left" rowspan="1" colspan="1">F 5′-TTCCAGCTGGCCCCTTGAGC-3′</td><td align="center" rowspan="2" colspan="1">146</td><td align="center" rowspan="2" colspan="1">1.873</td><td align="center" rowspan="2" colspan="1">0.982</td><td align="center" rowspan="2" colspan="1">[<xref rid="B53" ref-type="bibr">30</xref>]</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-GAGGGCATTGGCATACCCAC-3′</td></tr><tr><td align="center" colspan="7" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="2" colspan="1">COX-2</td><td align="left" rowspan="2" colspan="1">NM_214321</td><td align="left" rowspan="1" colspan="1">F 5′-AGAAGCGAGGACCAGCTTTC-3′</td><td align="center" rowspan="2" colspan="1">215</td><td align="center" rowspan="2" colspan="1">1.905</td><td align="center" rowspan="2" colspan="1">0.981</td><td align="center" rowspan="2" colspan="1">NCBI/Primer-Blast</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-AAAGCGGAGGTGTTCAGGAG-3′</td></tr><tr><td align="center" colspan="7" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="2" colspan="1">Hsp70</td><td align="left" rowspan="2" colspan="1">NM_001123127</td><td align="left" rowspan="1" colspan="1">F 5′-GCCCTGAATCCGCAGAATA-3′</td><td align="center" rowspan="2" colspan="1">152</td><td align="center" rowspan="2" colspan="1">2.0</td><td align="center" rowspan="2" colspan="1">0.993</td><td align="center" rowspan="2" colspan="1">[<xref rid="B54" ref-type="bibr">31</xref>]</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-TCCCCACGGTAGGAAACG-3′</td></tr><tr><td align="center" colspan="7" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="2" colspan="1">CycA</td><td align="left" rowspan="2" colspan="1">NM_214353</td><td align="left" rowspan="1" colspan="1">F 5′-GCGTCTCCTTCGAGCTGTT-3′</td><td align="center" rowspan="2" colspan="1">160</td><td align="center" rowspan="2" colspan="1">1.907</td><td align="center" rowspan="2" colspan="1">0.998</td><td align="center" rowspan="2" colspan="1">[<xref rid="B53" ref-type="bibr">30</xref>]</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-CCATTATGGCGTGTGAAGTC-3′</td></tr><tr><td align="center" colspan="7" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="2" colspan="1">HPRT</td><td align="left" rowspan="2" colspan="1">NM_001032376</td><td align="left" rowspan="1" colspan="1">F 5′-GGACTTGAATCATGTTTGTG-3′</td><td align="center" rowspan="2" colspan="1">91</td><td align="center" rowspan="2" colspan="1">1.963</td><td align="center" rowspan="2" colspan="1">0.997</td><td align="center" rowspan="2" colspan="1">[<xref rid="B55" ref-type="bibr">32</xref>]</td></tr><tr><td align="left" rowspan="1" colspan="1">R 5′-CAGATGTTTCCAAACTCAAC-3′</td></tr></tbody></table></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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