Induction of a 5-lipoxygenase product by daidzein is involved in the regulation of influenza virus replication
Identifieur interne : 000662 ( Pmc/Corpus ); précédent : 000661; suivant : 000663Induction of a 5-lipoxygenase product by daidzein is involved in the regulation of influenza virus replication
Auteurs : Yuka Horio ; Riho Sogabe ; Mototada Shichiri ; Noriko Ishida ; Ryosuke Morimoto ; Atsushi Ohshima ; Yuji IsegawaSource :
- Journal of Clinical Biochemistry and Nutrition [ 0912-0009 ] ; 2020.
Abstract
This study was conducted to evaluate the regulation mechanism of influenza virus replication following treatment of Madin-Darby canine kidney cells with the soy isoflavone daidzein. We performed comparative qualitative and quantitative analyses of lipid peroxide between mock-infected and virus-infected cells treated with or without daidzein, as it had been reported that daidzein was an antioxidant and lipid peroxide levels increased upon virus infection. Contrary to our belief, lipid peroxides were not elevated in virus-infected cells and no decrease in lipid peroxides was observed in daidzein-treated cells. In daidzein-treated cells, 5-hydroxyeicosatetraenoic acid, the 5-lipoxygenase product derived from arachidonate, was significantly elevated compared to other lipid peroxides. Zileuton (5-lipoxygenase inhibitor) and 5-lipoxygenase knockdown reduced the daidzein-induced antiviral effect. Moreover, virus replication was regulated by treatment with 5-hydroperoxyeicosatetraenoic acid, a precursor of 5-hydroxyeicosatetraenoic acid and 5-lipoxygenase primary product. These results suggest that daidzein regulates virus replication via signal transduction through 5-lipoxygenase products.
Url:
DOI: 10.3164/jcbn.19-70
PubMed: 32001954
PubMed Central: 6983437
Links to Exploration step
PMC:6983437Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Induction of a 5-lipoxygenase product by daidzein is involved in the regulation of influenza virus replication</title><author><name sortKey="Horio, Yuka" sort="Horio, Yuka" uniqKey="Horio Y" first="Yuka" last="Horio">Yuka Horio</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author><author><name sortKey="Sogabe, Riho" sort="Sogabe, Riho" uniqKey="Sogabe R" first="Riho" last="Sogabe">Riho Sogabe</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author><author><name sortKey="Shichiri, Mototada" sort="Shichiri, Mototada" uniqKey="Shichiri M" first="Mototada" last="Shichiri">Mototada Shichiri</name><affiliation><nlm:aff id="aff2">Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), 1-1-1 Higashi, Tsukuba-shi, Ibaraki 305-8562, Japan</nlm:aff></affiliation></author><author><name sortKey="Ishida, Noriko" sort="Ishida, Noriko" uniqKey="Ishida N" first="Noriko" last="Ishida">Noriko Ishida</name><affiliation><nlm:aff id="aff2">Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan</nlm:aff></affiliation></author><author><name sortKey="Morimoto, Ryosuke" sort="Morimoto, Ryosuke" uniqKey="Morimoto R" first="Ryosuke" last="Morimoto">Ryosuke Morimoto</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author><author><name sortKey="Ohshima, Atsushi" sort="Ohshima, Atsushi" uniqKey="Ohshima A" first="Atsushi" last="Ohshima">Atsushi Ohshima</name><affiliation><nlm:aff id="aff4">Genomics Program, Nagahama Institute of Bio-Science and Technology, 1266 Tamura-Cho, Nagahama, Shiga 526-0829, Japan</nlm:aff></affiliation></author><author><name sortKey="Isegawa, Yuji" sort="Isegawa, Yuji" uniqKey="Isegawa Y" first="Yuji" last="Isegawa">Yuji Isegawa</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation><affiliation><nlm:aff id="aff5">Institute for Biosciences, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">32001954</idno><idno type="pmc">6983437</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6983437</idno><idno type="RBID">PMC:6983437</idno><idno type="doi">10.3164/jcbn.19-70</idno><date when="2020">2020</date><idno type="wicri:Area/Pmc/Corpus">000662</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000662</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Induction of a 5-lipoxygenase product by daidzein is involved in the regulation of influenza virus replication</title><author><name sortKey="Horio, Yuka" sort="Horio, Yuka" uniqKey="Horio Y" first="Yuka" last="Horio">Yuka Horio</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author><author><name sortKey="Sogabe, Riho" sort="Sogabe, Riho" uniqKey="Sogabe R" first="Riho" last="Sogabe">Riho Sogabe</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author><author><name sortKey="Shichiri, Mototada" sort="Shichiri, Mototada" uniqKey="Shichiri M" first="Mototada" last="Shichiri">Mototada Shichiri</name><affiliation><nlm:aff id="aff2">Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), 1-1-1 Higashi, Tsukuba-shi, Ibaraki 305-8562, Japan</nlm:aff></affiliation></author><author><name sortKey="Ishida, Noriko" sort="Ishida, Noriko" uniqKey="Ishida N" first="Noriko" last="Ishida">Noriko Ishida</name><affiliation><nlm:aff id="aff2">Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan</nlm:aff></affiliation></author><author><name sortKey="Morimoto, Ryosuke" sort="Morimoto, Ryosuke" uniqKey="Morimoto R" first="Ryosuke" last="Morimoto">Ryosuke Morimoto</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author><author><name sortKey="Ohshima, Atsushi" sort="Ohshima, Atsushi" uniqKey="Ohshima A" first="Atsushi" last="Ohshima">Atsushi Ohshima</name><affiliation><nlm:aff id="aff4">Genomics Program, Nagahama Institute of Bio-Science and Technology, 1266 Tamura-Cho, Nagahama, Shiga 526-0829, Japan</nlm:aff></affiliation></author><author><name sortKey="Isegawa, Yuji" sort="Isegawa, Yuji" uniqKey="Isegawa Y" first="Yuji" last="Isegawa">Yuji Isegawa</name><affiliation><nlm:aff id="aff1">Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation><affiliation><nlm:aff id="aff5">Institute for Biosciences, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Clinical Biochemistry and Nutrition</title><idno type="ISSN">0912-0009</idno><idno type="eISSN">1880-5086</idno><imprint><date when="2020">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>This study was conducted to evaluate the regulation mechanism of influenza virus replication following treatment of Madin-Darby canine kidney cells with the soy isoflavone daidzein. We performed comparative qualitative and quantitative analyses of lipid peroxide between mock-infected and virus-infected cells treated with or without daidzein, as it had been reported that daidzein was an antioxidant and lipid peroxide levels increased upon virus infection. Contrary to our belief, lipid peroxides were not elevated in virus-infected cells and no decrease in lipid peroxides was observed in daidzein-treated cells. In daidzein-treated cells, 5-hydroxyeicosatetraenoic acid, the 5-lipoxygenase product derived from arachidonate, was significantly elevated compared to other lipid peroxides. Zileuton (5-lipoxygenase inhibitor) and 5-lipoxygenase knockdown reduced the daidzein-induced antiviral effect. Moreover, virus replication was regulated by treatment with 5-hydroperoxyeicosatetraenoic acid, a precursor of 5-hydroxyeicosatetraenoic acid and 5-lipoxygenase primary product. These results suggest that daidzein regulates virus replication via signal transduction through 5-lipoxygenase products.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Belser, Ja" uniqKey="Belser J">JA Belser</name></author><author><name sortKey="Maines, Tr" uniqKey="Maines T">TR Maines</name></author><author><name sortKey="Tumpey, Tm" uniqKey="Tumpey T">TM Tumpey</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chow, Ej" uniqKey="Chow E">EJ Chow</name></author><author><name sortKey="Doyle, Jd" uniqKey="Doyle J">JD Doyle</name></author><author><name sortKey="Uyeki, Tm" uniqKey="Uyeki T">TM Uyeki</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Le, Qm" uniqKey="Le Q">QM Le</name></author><author><name sortKey="Kiso, M" uniqKey="Kiso M">M Kiso</name></author><author><name sortKey="Someya, K" uniqKey="Someya K">K Someya</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bloom, Jd" uniqKey="Bloom J">JD Bloom</name></author><author><name sortKey="Gong, Li" uniqKey="Gong L">LI Gong</name></author><author><name sortKey="Baltimore, D" uniqKey="Baltimore D">D Baltimore</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Furuta, Y" uniqKey="Furuta Y">Y Furuta</name></author><author><name sortKey="Takahashi, K" uniqKey="Takahashi K">K Takahashi</name></author><author><name sortKey="Fukuda, Y" uniqKey="Fukuda Y">Y Fukuda</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kiso, M" uniqKey="Kiso M">M Kiso</name></author><author><name sortKey="Takahashi, K" uniqKey="Takahashi K">K Takahashi</name></author><author><name sortKey="Sakai Tagawa, Y" uniqKey="Sakai Tagawa Y">Y Sakai-Tagawa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nagai, E" uniqKey="Nagai E">E Nagai</name></author><author><name sortKey="Iwai, M" uniqKey="Iwai M">M Iwai</name></author><author><name sortKey="Koketsu, R" uniqKey="Koketsu R">R Koketsu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Christen, S" uniqKey="Christen S">S Christen</name></author><author><name sortKey="Peterhans, E" uniqKey="Peterhans E">E Peterhans</name></author><author><name sortKey="Stocker, R" uniqKey="Stocker R">R Stocker</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jacoby, Db" uniqKey="Jacoby D">DB Jacoby</name></author><author><name sortKey="Choi, Am" uniqKey="Choi A">AM Choi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Akaike, T" uniqKey="Akaike T">T Akaike</name></author><author><name sortKey="Noguchi, Y" uniqKey="Noguchi Y">Y Noguchi</name></author><author><name sortKey="Ijiri, S" uniqKey="Ijiri S">S Ijiri</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Jung, Ki" uniqKey="Jung K">KI Jung</name></author><author><name sortKey="Pyo, Cw" uniqKey="Pyo C">CW Pyo</name></author><author><name sortKey="Choi, Sy" uniqKey="Choi S">SY Choi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Franke, Aa" uniqKey="Franke A">AA Franke</name></author><author><name sortKey="Lai, Jf" uniqKey="Lai J">JF Lai</name></author><author><name sortKey="Halm, Bm" uniqKey="Halm B">BM Halm</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sankar, P" uniqKey="Sankar P">P Sankar</name></author><author><name sortKey="Zachariah, B" uniqKey="Zachariah B">B Zachariah</name></author><author><name sortKey="Vickneshwaran, V" uniqKey="Vickneshwaran V">V Vickneshwaran</name></author><author><name sortKey="Jacob, Se" uniqKey="Jacob S">SE Jacob</name></author><author><name sortKey="Sridhar, Mg" uniqKey="Sridhar M">MG Sridhar</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chen, W" uniqKey="Chen W">W Chen</name></author><author><name sortKey="Ma, X" uniqKey="Ma X">X Ma</name></author><author><name sortKey="Lin, Y" uniqKey="Lin Y">Y Lin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jin, M" uniqKey="Jin M">M Jin</name></author><author><name sortKey="Shen, Mh" uniqKey="Shen M">MH Shen</name></author><author><name sortKey="Jin, Mh" uniqKey="Jin M">MH Jin</name></author><author><name sortKey="Jin, Ah" uniqKey="Jin A">AH Jin</name></author><author><name sortKey="Yin, Xz" uniqKey="Yin X">XZ Yin</name></author><author><name sortKey="Quan, Js" uniqKey="Quan J">JS Quan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kuiper, Gg" uniqKey="Kuiper G">GG Kuiper</name></author><author><name sortKey="Carlsson, B" uniqKey="Carlsson B">B Carlsson</name></author><author><name sortKey="Grandien, K" uniqKey="Grandien K">K Grandien</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kuiper, Gg" uniqKey="Kuiper G">GG Kuiper</name></author><author><name sortKey="Lemmen, Jg" uniqKey="Lemmen J">JG Lemmen</name></author><author><name sortKey="Carlsson, B" uniqKey="Carlsson B">B Carlsson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Uif Lean, A" uniqKey="Uif Lean A">A Uifälean</name></author><author><name sortKey="Schneider, S" uniqKey="Schneider S">S Schneider</name></author><author><name sortKey="Ionescu, C" uniqKey="Ionescu C">C Ionescu</name></author><author><name sortKey="Lalk, M" uniqKey="Lalk M">M Lalk</name></author><author><name sortKey="Iuga, Ca" uniqKey="Iuga C">CA Iuga</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Folman, Y" uniqKey="Folman Y">Y Folman</name></author><author><name sortKey="Pope, Gs" uniqKey="Pope G">GS Pope</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yan, Gr" uniqKey="Yan G">GR Yan</name></author><author><name sortKey="Xiao, Cl" uniqKey="Xiao C">CL Xiao</name></author><author><name sortKey="He, Gw" uniqKey="He G">GW He</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hafez, Hs" uniqKey="Hafez H">HS Hafez</name></author><author><name sortKey="Ghareeb, Da" uniqKey="Ghareeb D">DA Ghareeb</name></author><author><name sortKey="Saleh, Sr" uniqKey="Saleh S">SR Saleh</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kim, Ms" uniqKey="Kim M">MS Kim</name></author><author><name sortKey="Kim, B" uniqKey="Kim B">B Kim</name></author><author><name sortKey="Park, H" uniqKey="Park H">H Park</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yoshida, Y" uniqKey="Yoshida Y">Y Yoshida</name></author><author><name sortKey="Kodai, S" uniqKey="Kodai S">S Kodai</name></author><author><name sortKey="Takemura, S" uniqKey="Takemura S">S Takemura</name></author><author><name sortKey="Minamiyama, Y" uniqKey="Minamiyama Y">Y Minamiyama</name></author><author><name sortKey="Niki, E" uniqKey="Niki E">E Niki</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Umeno, A" uniqKey="Umeno A">A Umeno</name></author><author><name sortKey="Shichiri, M" uniqKey="Shichiri M">M Shichiri</name></author><author><name sortKey="Ishida, N" uniqKey="Ishida N">N Ishida</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shichiri, M" uniqKey="Shichiri M">M Shichiri</name></author><author><name sortKey="Ishida, N" uniqKey="Ishida N">N Ishida</name></author><author><name sortKey="Hagihara, Y" uniqKey="Hagihara Y">Y Hagihara</name></author><author><name sortKey="Yoshida, Y" uniqKey="Yoshida Y">Y Yoshida</name></author><author><name sortKey="Kume, A" uniqKey="Kume A">A Kume</name></author><author><name sortKey="Suzuki, H" uniqKey="Suzuki H">H Suzuki</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lisovyy, Oo" uniqKey="Lisovyy O">OO Lisovyy</name></author><author><name sortKey="Dosenko, Ve" uniqKey="Dosenko V">VE Dosenko</name></author><author><name sortKey="Nagibin, Vs" uniqKey="Nagibin V">VS Nagibin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shichiri, M" uniqKey="Shichiri M">M Shichiri</name></author><author><name sortKey="Kono, N" uniqKey="Kono N">N Kono</name></author><author><name sortKey="Shimanaka, Y" uniqKey="Shimanaka Y">Y Shimanaka</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Steinhilber, D" uniqKey="Steinhilber D">D Steinhilber</name></author><author><name sortKey="Fischer, As" uniqKey="Fischer A">AS Fischer</name></author><author><name sortKey="Metzner, J" uniqKey="Metzner J">J Metzner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Liu, H" uniqKey="Liu H">H Liu</name></author><author><name sortKey="Zhong, H" uniqKey="Zhong H">H Zhong</name></author><author><name sortKey="Leng, L" uniqKey="Leng L">L Leng</name></author><author><name sortKey="Jiang, Z" uniqKey="Jiang Z">Z Jiang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="B Ck, M" uniqKey="B Ck M">M Bäck</name></author><author><name sortKey="Yurdagul, A" uniqKey="Yurdagul A">A Yurdagul</name></author><author><name sortKey="Tabas, I" uniqKey="Tabas I">I Tabas</name></author><author><name sortKey="Oorni, K" uniqKey="Oorni K">K Öörni</name></author><author><name sortKey="Kovanen, Pt" uniqKey="Kovanen P">PT Kovanen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Serhan, Cn" uniqKey="Serhan C">CN Serhan</name></author><author><name sortKey="Dalli, J" uniqKey="Dalli J">J Dalli</name></author><author><name sortKey="Colas, Ra" uniqKey="Colas R">RA Colas</name></author><author><name sortKey="Winkler, Jw" uniqKey="Winkler J">JW Winkler</name></author><author><name sortKey="Chiang, N" uniqKey="Chiang N">N Chiang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Morita, M" uniqKey="Morita M">M Morita</name></author><author><name sortKey="Kuba, K" uniqKey="Kuba K">K Kuba</name></author><author><name sortKey="Ichikawa, A" uniqKey="Ichikawa A">A Ichikawa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="R Dmark, O" uniqKey="R Dmark O">O Rådmark</name></author><author><name sortKey="Werz, O" uniqKey="Werz O">O Werz</name></author><author><name sortKey="Steinhilber, D" uniqKey="Steinhilber D">D Steinhilber</name></author><author><name sortKey="Samuelsson, B" uniqKey="Samuelsson B">B Samuelsson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Recchiuti, A" uniqKey="Recchiuti A">A Recchiuti</name></author><author><name sortKey="Serhan, Cn" uniqKey="Serhan C">CN Serhan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="King, Ra" uniqKey="King R">RA King</name></author><author><name sortKey="Bursill, Db" uniqKey="Bursill D">DB Bursill</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cheng, Wc" uniqKey="Cheng W">WC Cheng</name></author><author><name sortKey="Lo, Sc" uniqKey="Lo S">SC Lo</name></author><author><name sortKey="Tsai, Ks" uniqKey="Tsai K">KS Tsai</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Clin Biochem Nutr</journal-id><journal-id journal-id-type="iso-abbrev">J Clin Biochem Nutr</journal-id><journal-id journal-id-type="publisher-id">JCBN</journal-id><journal-title-group><journal-title>Journal of Clinical Biochemistry and Nutrition</journal-title></journal-title-group><issn pub-type="ppub">0912-0009</issn><issn pub-type="epub">1880-5086</issn><publisher><publisher-name>the Society for Free Radical Research Japan</publisher-name><publisher-loc>Kyoto, Japan</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">32001954</article-id><article-id pub-id-type="pmc">6983437</article-id><article-id pub-id-type="publisher-id">DN/JST.JSTAGE/jcbn/19-70</article-id><article-id pub-id-type="doi">10.3164/jcbn.19-70</article-id><article-id pub-id-type="publisher-manuscript">19-70</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>Induction of a 5-lipoxygenase product by daidzein is involved in the regulation of influenza virus replication</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Horio</surname><given-names>Yuka</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="author-notes" rid="fn1">†</xref></contrib><contrib contrib-type="author"><name><surname>Sogabe</surname><given-names>Riho</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="author-notes" rid="fn1">†</xref></contrib><contrib contrib-type="author"><name><surname>Shichiri</surname><given-names>Mototada</given-names></name><xref ref-type="aff" rid="aff2">2</xref><xref ref-type="aff" rid="aff3">3</xref><xref ref-type="author-notes" rid="fn1">†</xref></contrib><contrib contrib-type="author"><name><surname>Ishida</surname><given-names>Noriko</given-names></name><xref ref-type="aff" rid="aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Morimoto</surname><given-names>Ryosuke</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Ohshima</surname><given-names>Atsushi</given-names></name><xref ref-type="aff" rid="aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Isegawa</surname><given-names>Yuji</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="aff" rid="aff5">5</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib><aff id="aff1"><label>1</label>Department of Food Sciences and Nutrition, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</aff><aff id="aff2"><label>2</label>Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan</aff><aff id="aff3"><label>3</label>DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), 1-1-1 Higashi, Tsukuba-shi, Ibaraki 305-8562, Japan</aff><aff id="aff4"><label>4</label>Genomics Program, Nagahama Institute of Bio-Science and Technology, 1266 Tamura-Cho, Nagahama, Shiga 526-0829, Japan</aff><aff id="aff5"><label>5</label>Institute for Biosciences, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo 663-8558, Japan</aff></contrib-group><author-notes><fn id="fn1"><label>†</label><p>These authors contributed equally: YH, RS, MS.</p></fn><corresp id="cor1">*To whom correspondence should be addressed. E-mail: <email>isegawa@mukogawa-u.ac.jp</email></corresp></author-notes><pub-date pub-type="ppub"><month>1</month><year>2020</year></pub-date><pub-date pub-type="epub"><day>1</day><month>1</month><year>2020</year></pub-date><volume>66</volume><issue>1</issue><fpage>36</fpage><lpage>42</lpage><history><date date-type="received"><day>17</day><month>8</month><year>2019</year></date><date date-type="accepted"><day>29</day><month>9</month><year>2019</year></date></history><permissions><copyright-statement>Copyright © 2020 JCBN</copyright-statement><copyright-year>2020</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/3.0/"><license-p>This is an open access article distributed under the terms of 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>This study was conducted to evaluate the regulation mechanism of influenza virus replication following treatment of Madin-Darby canine kidney cells with the soy isoflavone daidzein. We performed comparative qualitative and quantitative analyses of lipid peroxide between mock-infected and virus-infected cells treated with or without daidzein, as it had been reported that daidzein was an antioxidant and lipid peroxide levels increased upon virus infection. Contrary to our belief, lipid peroxides were not elevated in virus-infected cells and no decrease in lipid peroxides was observed in daidzein-treated cells. In daidzein-treated cells, 5-hydroxyeicosatetraenoic acid, the 5-lipoxygenase product derived from arachidonate, was significantly elevated compared to other lipid peroxides. Zileuton (5-lipoxygenase inhibitor) and 5-lipoxygenase knockdown reduced the daidzein-induced antiviral effect. Moreover, virus replication was regulated by treatment with 5-hydroperoxyeicosatetraenoic acid, a precursor of 5-hydroxyeicosatetraenoic acid and 5-lipoxygenase primary product. These results suggest that daidzein regulates virus replication via signal transduction through 5-lipoxygenase products.</p></abstract><kwd-group kwd-group-type="author"><kwd>influenza virus</kwd><kwd>daidzein</kwd><kwd>lipid peroxide</kwd><kwd>signal transduction</kwd><kwd>lipoxygenase</kwd></kwd-group></article-meta></front><body><sec sec-type="introduction"><title>Introduction</title><p>Influenza viruses, particularly A viruses, cause epidemics and pandemics in human populations, resulting in enormous suffering and economic losses.<sup>(<xref rid="B1" ref-type="bibr">1</xref>)</sup> Influenza virus infections can cause severe complications such as pneumonia and encephalitis and can increase the rate of hospitalization and mortality, particularly in young children and elderly people.<sup>(<xref rid="B2" ref-type="bibr">2</xref>,<xref rid="B3" ref-type="bibr">3</xref>)</sup> The M2 ion channel inhibitors amantadine and rimantadine and neuraminidase inhibitors zanamivir and oseltamivir have been used to treat influenza virus infections. Recently, neuraminidase inhibitor-resistant influenza A viruses were reported and an antiviral reagent against RNA-dependent RNA polymerase in influenza virus was developed.<sup>(<xref rid="B4" ref-type="bibr">4</xref>–<xref rid="B7" ref-type="bibr">7</xref>)</sup></p><p>We previously reported that soybean extract exerted antiviral activity against influenza virus growth.<sup>(<xref rid="B8" ref-type="bibr">8</xref>)</sup> During influenza infection, reactive oxygen species (ROS) and lipid peroxides are generated in several tissues such as the lung.<sup>(<xref rid="B9" ref-type="bibr">9</xref>–<xref rid="B13" ref-type="bibr">13</xref>)</sup> Soy isoflavones, such as daidzein, glycitein, and genistein,<sup>(<xref rid="B14" ref-type="bibr">14</xref>)</sup> are a group of compounds present in soybean and function as potent antioxidants.<sup>(<xref rid="B15" ref-type="bibr">15</xref>,<xref rid="B16" ref-type="bibr">16</xref>)</sup> Jin <italic>et al.</italic><sup>(<xref rid="B17" ref-type="bibr">17</xref>)</sup> reported the effect of soy isoflavones, including daidzein (61.7%), glycitein (21.5%) and genistein (16.2%), on diabetic rats. They showed that an improvement in postprandial blood glucose levels and a significant suppression of blood lipid peroxide can be achieved by single or long-term administration of soy isoflavones, respectively.<sup>(<xref rid="B17" ref-type="bibr">17</xref>)</sup> Additionally, soy isoflavones act as phytoestrogens and interact with animal and human estrogen receptors and produce non-hormonal effects.<sup>(<xref rid="B18" ref-type="bibr">18</xref>–<xref rid="B24" ref-type="bibr">24</xref>)</sup> We found that daidzein and glycitein inhibited influenza virus replication and suggested that the regulation by daidzein is not directly associated with viral enzymes but rather with host-cellular metabolism (unpublished data).</p><p>In this study, we first confirmed whether daidzein exerts antiviral activity through its antioxidant function by measuring lipid peroxide levels. Furthermore, we identified a lipid oxidation product specifically increased in daidzein-treated cells as a lipid mediator that induces antiviral activity and analyzed its production mechanism.</p></sec><sec sec-type="materials and methods"><title>Materials and Methods</title><sec><title>Cells and viruses</title><p>Madin-Darby canine kidney (MDCK) cells were grown in Eagle’s minimum essential medium (MEM; Sigma-Aldrich, St. Louis, MO) containing 70 ml/L fetal bovine serum (FBS). The influenza A virus H1N1 (PR/8/34) was used throughout the experiments. For cell infection, the virus was diluted in serum-free MEM supplemented with 0.4 g/L bovine serum albumin (Sigma-Aldrich), which was adsorbed to cells at a multiplicity of infection (MOI) of 0.001 for 1 h at 37°C. The inoculum then was removed and replaced with FBS-free Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 4 g/L bovine serum albumin and acetyltrypsin (2 µg/mL; Sigma-Aldrich) for the remainder of the infection period.</p></sec><sec><title>Focus forming reduction assay for virucidal activity</title><p>Sample influenza viruses used in the virucidal activity assay were prepared by infecting monolayer sheets of MDCK cells with viruses (100 µl, MOI 0.001) in 24-well plates for 1 h as described above. The inoculum was removed, washed once with serum-free MEM, and FBS-free DMEM supplemented with daidzein was added (Kanto Chemical Co., Inc., Tokyo, Japan). After 24 h, the supernatants were harvested and used as the influenza virus samples for focus forming reduction assay (FFRA).</p><p>Focus formation was induced as described by Nagai <italic>et al.</italic><sup>(<xref rid="B8" ref-type="bibr">8</xref>)</sup> Each virus dilution was selected to give a final count of approximately 30 focus-forming units (FFU) per well. Virucidal activity was expressed as the reciprocal of the highest dilution that reduced the number of foci to ≤50% of the control value.</p></sec><sec><title>Sample preparation for lipid peroxide</title><p>Virus-infected cells for analysis of lipid peroxidation products were prepared by infecting monolayer sheets of MDCK cells (1 ml, MOI 0.001) in 10-cm dishes for 1 h as described above. The inoculum then was removed and washed once with serum-free MEM, after which FBS-free DMEM supplemented with or without 275 µM daidzein was added. After 24 h, the dishes were washed twice with phosphate-buffered saline (PBS). The cells were harvested into 1.5-ml tubes containing 500 µl of ice-cold PBS using a cell-scraper. The samples were placed on ice immediately after collection. The cells were obtained by centrifugation at 12,500 × <italic>g</italic> for 5 min at 4°C, the supernatant was discarded, and 140 µl of ice-cold PBS was added to the tube. The cells were homogenized with a sonicator. The homogenate (110 µl) was transferred to a new tube, and then 4 volumes of methanol containing 100 µM 2,6-di-<italic>t</italic>-butyl-4-methylphenol (Wako, Osaka, Japan) were added. The tube was mixed by vortexing for 1 min, followed by centrifugation at 12,500 × <italic>g</italic> for 5 min at 4°C. The supernatant (500 µl) was either stored at −80°C or analyzed immediately to detect lipid peroxidation products. Lipid peroxide contents were normalized to the protein concentration, which was measured by BCA protein assay.</p></sec><sec><title>Analysis of lipid peroxides</title><p>To analyze lipid peroxides, the concentration of 7β-hydroxycholesterol (7β-OHCh), a cholesterol-derived peroxidation product, 4 isomers of hydroxyoctadecadienoic acid (HODE), which are linoleate-derived peroxidation products, and 3 isomers of hydroxyeicosatetraenoic acid (HETE) and 8-iso-prostaglandin F<sub>2α</sub> (8-iso-PGF<sub>2α</sub>), which are arachidonate-derived peroxidation products, were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (HODEs, HETEs, and 8-iso-PGF<sub>2α</sub>) or gas chromatography-mass spectrometry (7β-OHCh) as previously described.<sup>(<xref rid="B25" ref-type="bibr">25</xref>–<xref rid="B27" ref-type="bibr">27</xref>)</sup> 13-Hydroxy-9(<italic>E</italic>), 11(<italic>E</italic>)-octadecadienoic acid [13-(<italic>E</italic>,<italic>E</italic>)-HODE], 9-hydroxy-10(<italic>E</italic>), 12(<italic>E</italic>)-octadecadienoic acid [9-(<italic>E</italic>,<italic>E</italic>)-HODE], 7β-OHCh, and 8-iso-PGF<sub>2α</sub> are specific products of radical-mediated oxidation. 13-Hydroxy-9(<italic>Z</italic>),11(<italic>E</italic>)-octadecadienoic acid [13-(<italic>Z</italic>,<italic>E</italic>)-HODE], and 5-, 12-, and 15-HETE are generated by both radical-mediated oxidation and enzymatic oxidation. Total HODE (tHODE) indicates the sum of the 4 isomers of HODEs.</p></sec><sec><title>Knockdown of 5-lipoxygenase in MDCK cells</title><p>For RNA interference, sequences were produced as described by Lisovyy <italic>et al.</italic><sup>(<xref rid="B28" ref-type="bibr">28</xref>)</sup> A 5-lipoxygenase (5-LOX)-specific short interfering RNA (siRNA) (sense 5'-GCAAGAAGCUACCCGAGAAUU-3', and antisense 5'-UUCUCGGGUAGCUUCUUGCUU-3') was prepared from corresponding oligonucleotides provided by Dharmacon (Lafayette, CO) according to the manufacturer’s protocol. MDCK cells were transfected with siRNA (5 nM final concentration) using lipofectamine RNAiMAX (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. At 24 h after siRNA transfection, the cells were placed in fresh medium. At 48 h after siRNA transfection, viruses (MOI 0.001) were infected for 1 h, and the inoculum was removed, washed once with serum-free MEM, and replaced with FBS-free DMEM supplemented with or without 275 µM daidzein.</p></sec><sec><title>Western blot analysis of 5-LOX protein</title><p>Whole cell lysates of MDCK cells were prepared by homogenization in RIPA buffer. The whole cell lysate of MDCK cells was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 5-LOX and actin were analyzed by immunoblotting using an anti-5-LOX monoclonal antibody (ab169755, dilution 1/1,000; Abcam, Cambridge, UK) and anti-actin antibody (MAB1501R, dilution 1/2,000; Chemicon International, Temecula, CA).</p></sec><sec><title>Evaluation of LOX activities in MDCK cell lysates</title><p> The cells for analysis of 5-LOX activity were prepared by culturing non-infected monolayer sheets of MDCK cells in 10-cm dishes supplemented with or without 275 µM daidzein (mock-treated cells: 3 dishes and daidzein-treated cells: 3 dishes). After 24 h, the dishes were washed twice with PBS. The cells were harvested into 1.5-ml tubes containing 500 µl of 2% bovine serum albumin containing DMEM using a cell-scraper. The cells were homogenized and then evenly divided into six tubes. Arachidonic acid (AA) was added to each tube to a final concentration of 120 µM and incubated for 180 min at 37°C with gentle mixing with a microtube mixer. After 180 min of incubation, 4 volumes of methanol containing 100 µM 2,6-di-<italic>t</italic>-butyl-4-methylphenol (Wako) was added. The tubes were mixed by vortexing for 1 min, and then centrifuged at 12,500 × <italic>g</italic> for 5 min at 4°C. The supernatant (500 µl) was either stored at −80°C or used to detect 5-, 12-, and 15-HETE and AA. HETEs and AA were measured as described above for lipid peroxide analysis. The content of HETEs and AA was normalized to the protein concentration, which was measured by BCA protein assay.</p></sec><sec><title>Cytotoxicity analysis</title><p>The WST-8 assay [using Cell Counting Kit-8 (Dojindo Laboratories, Kumamoto, Japan)] is a modified MTT assay that measures the mitochondrial reduction capacity and can quantify cell viability.<sup>(<xref rid="B29" ref-type="bibr">29</xref>)</sup> After treatment with daidzein or 5-hydroperoxyeicosatetraenoic acid (5-HpETE), the cells in 96-well plates were incubated with 10 µl of Cell Counting Kit-8 solution {containing with 4-[3-(2-methoxy-4-nitrophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate sodium salt} in medium for 1 h at 37°C. Absorbance was measured photometrically at 450 nm. Cell viabilities were expressed as a percentage of the absorbance measured in non-treated cells.</p></sec><sec><title>Statistical analysis</title><p>Statistical analyses were performed using unpaired <italic>t</italic> test and analysis of variance with Tukey-Kramer test using SPSS ver. 21.0 software (SPSS, Inc., Chicago, IL). A <italic>p</italic> value of less than 0.05 was considered to indicate significance.</p></sec></sec><sec sec-type="results"><title>Results</title><sec><title>Effect of virus replication and lipid peroxide production in MDCK cells by daidzein addition</title><p>The titer of influenza virus in the culture medium decreased in a dose-dependent manner following addition of daidzein (Fig. <xref ref-type="fig" rid="F1">1</xref>A). Daidzein inhibited influenza virus multiplication at an IC<sub>50</sub> of 51.2 µM (Fig. <xref ref-type="fig" rid="F1">1</xref>A). The effect of daidzein addition on the cell viability of MDCK cells was analyzed, however, it was found that cell proliferation was not affected by up to 400 µM of daidzein (Fig. <xref ref-type="fig" rid="F1">1</xref>B). To clarify the mechanism of antiviral activity of daidzein, the following experiments were performed with the addition of a daidzein concentration fixed at 275 µM. This concentration of daidzein in the culture medium inhibited influenza virus multiplication by approximately 85% and did not exert toxicity on MDCK cells.</p><p>The contents of lipid peroxide in the cells are shown in Fig. <xref ref-type="fig" rid="F2">2</xref>A–F. Lipid peroxide was not elevated in virus-infected cells (Fig. <xref ref-type="fig" rid="F2">2</xref>A–F). Although daidzein is known as an antioxidant, lipid peroxide was not decreased in daidzein-treated cells (Fig. <xref ref-type="fig" rid="F2">2</xref>A–F). These results indicate that daidzein did not exert antiviral activity through its antioxidant function under the conditions of this experiment. In contrast, 5-HETE content in the cells significantly increased in daidzein-treated cells both with and without virus infection (Fig. <xref ref-type="fig" rid="F2">2</xref>F), although the contents of 7β-OHCh (Fig. <xref ref-type="fig" rid="F2">2</xref>A), tHODE (Fig. <xref ref-type="fig" rid="F2">2</xref>B), 8-iso-PGF<sub>2α</sub> (Fig. <xref ref-type="fig" rid="F2">2</xref>C), 12-HETE (Fig. <xref ref-type="fig" rid="F2">2</xref>D), and 15-HETE (Fig. <xref ref-type="fig" rid="F2">2</xref>E) were not significantly increased in daidzein-treated cells.</p><p>As daidzein addition specifically induced a significant 5-HETE production in MDCK cells, we hypothesized that daidzein activated 5-lipoxygenase (5-LOX), which converts arachidonic acid to 5-HpETE, a precursor of 5-HETE. We analyzed the 5-, 12-, and 15-LOX activities in MDCK cell lysates. The cell lysates were obtained from mock-treated MDCK cells or those treated with daidzein and incubated with 120 µM arachidonic acid (AA). After 180 min incubation, 5-, 12-, and 15-HETE concentrations in the reactants were analyzed by LC-MS/MS. As shown in Fig. <xref ref-type="fig" rid="F3">3</xref>A, 5-HETE production in the daidzein-treated cell lysate was significantly higher than that in the mock-treated cell lysate. In contrast, daidzein treatment did not activate 12-HETE (Fig. <xref ref-type="fig" rid="F3">3</xref>B) and 15-HETE (Fig. <xref ref-type="fig" rid="F3">3</xref>C) production. These data indicate that daidzein enhances the enzymatic activity of 5-LOX.</p></sec><sec><title>Effect of zileuton on influenza virus replication in daidzein added virus-infected cells</title><p>To confirm that 5-LOX is involved in the daidzein-induced antiviral activity, we examined the effect of inhibitors of 5-LOX on antiviral activity. The antiviral activity of daidzein was significantly reduced following treatment with zileuton, a selective direct inhibitor of 5-LOX (Fig. <xref ref-type="fig" rid="F4">4</xref>A). Daidzein-induced inhibition of virus replication was decreased in the presence of zileuton from 72% [flu: flu + daidzein, inhibition rate = 100 – 100 × 0.58 × 10<sup>6</sup>/2.1 × 10<sup>6</sup> (%)] to 51% [flu + zileuton: flu + daidzein + zileuton, inhibition rate = 100 – 100 × 1.2 × 10<sup>6</sup>/2.5 × 10<sup>6</sup> (%)] (Fig. <xref ref-type="fig" rid="F4">4</xref>A). The virus titer was increased by zileuton to 212% [flu + daidzein: flu + daidzein + zileuton, activation rate = 100 × 1.2 × 10<sup>6</sup>/0.58 × 10<sup>6</sup> (%)] (Fig. <xref ref-type="fig" rid="F4">4</xref>A). Zileuton tended to reduce daidzein-induced 5-HETE production, but the reduction was not significant (Fig. <xref ref-type="fig" rid="F4">4</xref>B).</p><p>In contrast, when MK-886 was used to indirectly inhibit 5-LOX, daidzein-induced antiviral activity was not inhibited (Supplemental Fig. <xref ref-type="supplementary-material" rid="SF1">1</xref>A<bold>*</bold>). Additionally, MK-886 did not affect 5-HETE production in daidzein-treated cells (Supplemental Fig. <xref ref-type="supplementary-material" rid="SF1">1</xref>B). This suggests that MK-886 does not inhibit 5-LOX activity in MDCK cells.</p></sec><sec><title>Effect of 5-LOX knockdown on antiviral activity of daidzein</title><p>We hypothesized that daidzein affected 5-LOX-mediated enzymatic oxidation of AA, as zileuton inhibited daidzein-induced antiviral activity. We attempted to knockdown endogenous 5-LOX by transfection with a siRNA against canine-5-LOX. Based on the immunoblotting results (Fig. <xref ref-type="fig" rid="F4">4</xref>C), 5-LOX expression levels in MDCK cells were suppressed. We then measured the influenza virus titer in the culture medium. In control siRNA-transfected MDCK cells, daidzein inhibited virus replication by approximately 50.8% [flu: flu + daidzein, inhibition rate = 100 – 100 × 4.91 × 10<sup>6</sup>/9.99 × 10<sup>6</sup> (%)] (Fig. <xref ref-type="fig" rid="F4">4</xref>D). In contrast, following knockdown of 5-LOX, daidzein reduced virus replication to 26.5% [flu: flu + daidzein, inhibition rate = 100 – 100 × 7.07 × 10<sup>6</sup>/9.62 × 10<sup>6</sup> (%)] (Fig. <xref ref-type="fig" rid="F4">4</xref>D). The difference in the viral titer between daidzein-treated control siRNA MDCK cells (4.91 ± 1.18 × 10<sup>6</sup> FFU/ml) and daidzein-treated 5-LOX knockdown MDCK cells (7.07 ± 1.53 × 10<sup>6</sup> FFU/ml) was significant (<italic>p</italic><0.05). These results indicate that 5-LOX is involved in the inhibitory effect of daidzein on virus replication.</p></sec><sec><title>Effect of 5-HpETE administration on viral replication in virus-infected MDCK cells</title><p>Because 5-LOX was found to be involved in daidzein-induced inhibition of virus replication, we confirmed whether 5-LOX product inhibits virus proliferation. Administration of 5-HpETE, a precursor of 5-HETE and 5-LOX primary product derived from AA, inhibited virus proliferation in a concentration-dependent manner (Fig. <xref ref-type="fig" rid="F5">5</xref>A) with an IC<sub>50</sub> of 204.0 ng/ml (606.3 nM). 5-HpETE showed nearly no cytotoxicity at concentrations up to 1,250 ng/ml (Fig. <xref ref-type="fig" rid="F5">5</xref>B). These results indicate that 5-HpETE or its metabolite regulate virus replication.</p></sec></sec><sec sec-type="discussion"><title>Discussion</title><p>The present study clarified the mechanism of daidzein-induced inhibition of influenza virus replication. Daidzein showed no antioxidant activity under our experimental conditions (Fig. <xref ref-type="fig" rid="F2">2</xref>A–F); however, it enhanced 5-HETE production (Fig. <xref ref-type="fig" rid="F2">2</xref>F). Based on the effects of zileuton, a direct inhibitor of 5-LOX (Fig. <xref ref-type="fig" rid="F4">4</xref>A and B), and knockdown of 5-LOX (Fig. <xref ref-type="fig" rid="F4">4</xref>D), we observed that 5-LOX is involved in daidzein-induced 5-HETE production and daidzein-induced anti-influenza activity. Additionally, 5-HpETE, the primary product of 5-LOX, inhibited influenza virus replication (Fig. <xref ref-type="fig" rid="F5">5</xref>A).</p><p>This is the first study to demonstrate that daidzein activates 5-LOX and regulates viral replication. No previous studies have reported exogenous compounds that can increase 5-LOX activity, although 5-lipoxygenase-activating protein (FLAP) is well known as an endogenous 5-LOX activator.<sup>(<xref rid="B30" ref-type="bibr">30</xref>)</sup> The food component daidzein enhances 5-LOX activity and can be used to prevent and treat influenza. Liu <italic>et al.</italic><sup>(<xref rid="B31" ref-type="bibr">31</xref>)</sup> reported the effect of soy isoflavone administration to non-alcoholic fatty liver disease model rat. And they showed that soy isoflavones affected the expression of sterol regulatory element binding protein (SREBP)-1c and peroxisome proliferator-activated receptor (PPAR) α.<sup>(<xref rid="B31" ref-type="bibr">31</xref>)</sup> There may be potential signal transduction pathways to enhance 5-LOX activity by daidzein.</p><p>It has been reported that oxidation products derived from omega-3 and omega-6 polyunsaturated fatty acids (PUFAs) have various functions such as causing and resolving inflammation and are involved in signal transduction as lipid mediators.<sup>(<xref rid="B32" ref-type="bibr">32</xref>,<xref rid="B33" ref-type="bibr">33</xref>)</sup> Morita <italic>et al.</italic><sup>(<xref rid="B34" ref-type="bibr">34</xref>)</sup> reported that the omega-3 PUFA-derived lipid mediator protectin D1 (PD1) attenuates influenza virus replication. They performed a screen of PUFA-derived lipid metabolites in human lung epithelial (A549) cells infected with influenza A virus (PR8 virus) by using bioactive lipid libraries containing prostaglandins, resolvins, protectins, and other PUFA-derived metabolites.<sup>(<xref rid="B34" ref-type="bibr">34</xref>)</sup> They evaluated the antiviral effect by influenza virus nucleoprotein mRNA expression at 8 h after infection and found that 12-HETE, 15-HETE, 17-hydroxy-4<italic>Z</italic>,7<italic>Z</italic>,10<italic>Z</italic>,13<italic>Z</italic>,15<italic>E</italic>,19<italic>Z</italic>-docosahexaenoic acid, or PD1 treatment of PR8-virus-infected A549 cells inhibited nucleoprotein mRNA expression.<sup>(<xref rid="B34" ref-type="bibr">34</xref>)</sup> Results of their report indicate that there are several PUFA-derived metabolites, which have anti-influenza activity, produced by lipid-metabolizing enzymes such as lipoxygenase. We demonstrated the antiviral activity of 5-HpETE in the present study (Fig. <xref ref-type="fig" rid="F5">5</xref>A); however, the mechanism remains unclear. 5-HpETE is further metabolized to leukotrienes by leukotriene-converting enzymes and leukotrienes exert physiological functions in the pathophysiology of asthma and inflammation.<sup>(<xref rid="B35" ref-type="bibr">35</xref>)</sup> Additionally, there is another 5-LOX associated arachidonate-derived metabolite: lipoxin A4.<sup>(<xref rid="B36" ref-type="bibr">36</xref>)</sup> It is possible that daidzein-induced enzymatic activation of 5-LOX produced 5-LOX-associated PUFA-derived metabolites other than 5-HpETE and that these metabolites exert antiviral activity. Further studies are needed to determine whether 5-HpETE or another 5-LOX-associated PUFA-derived metabolite exerts antiviral functions and how these lipid metabolites suppress viral replication.</p><p>Zileuton, a direct inhibitor of 5-LOX, significantly reduced the antiviral activity of daidzein (Fig. <xref ref-type="fig" rid="F4">4</xref>A). However, the reduction effect of zileuton on daidzein-induced 5-HETE production was not significant (Fig. <xref ref-type="fig" rid="F4">4</xref>B). As shown in Fig. <xref ref-type="fig" rid="F5">5</xref>A, the inhibition rate of viral replication was elevated sharply at low concentrations of 5-HpETE (<200 ng/ml). This indicates that even a slight reduction in 5-HETE by zileuton addition can affect viral replication. Additionally, 5-HETE is produced in cells not only by 5-LOX-mediated enzymatic oxidation, but also by oxidation by ROS. Although we did not determine the proportion of 5-HETE generated by ROS relative to the total 5-HETE level, zileuton did not effectively suppress 5-HETE production because zileuton did not suppress ROS-mediated 5-HETE generation. This may be because zileuton could not significantly reduce 5-HETE generation without daidzein administration (Fig. <xref ref-type="fig" rid="F4">4</xref>B). As shown in Supplemental Fig. <xref ref-type="supplementary-material" rid="SF1">1</xref>, MK-886, an inhibitor of 5-LOX, did not suppress 5-HETE production. MK-886 inhibits FLAP, which is thought to facilitate the transfer of phospholipid-derived AA to 5-LOX.<sup>(<xref rid="B30" ref-type="bibr">30</xref>)</sup> It is thought that MK-886 cannot inactivate FLAP in MDCK cells.</p><p>The addition of daidzein showed anti-influenza activity with an IC<sub>50</sub> of 51.2 µM (Fig. <xref ref-type="fig" rid="F1">1</xref>A). King <italic>et al.</italic><sup>(<xref rid="B37" ref-type="bibr">37</xref>)</sup> reported the pharmacokinetics of the soy isoflavones daidzein and genistein in humans. Subjects consumed single soybean flour-based meal (0.84 g flour/kg body weight) and plasma concentration of isoflavones were analyzed throughout the 35-h post-meal period. This soybean flour-based meal provided 2.7 µmol/kg body weight of daidzein. Daidzein concentration in plasma reached maximum values of 3.14 ± 0.36 µmol/L.<sup>(<xref rid="B37" ref-type="bibr">37</xref>)</sup> A subject weighting 60 kg would have consumed 41.2 mg (162 µmol) of daidzein. Based on the results of this report, in order to increase the plasma concentration of daidzein to 51.2 µM, the IC<sub>50</sub> for anti-influenza activity, it is estimated that 667.4 mg of daidzein must be consumed. On the other hand, Cheng <italic>et al.</italic><sup>(<xref rid="B38" ref-type="bibr">38</xref>)</sup> reported the effect of a high dose of isoflavones (300 mg/day) on coagulation function in postmenopausal women. After 1-year treatment with a high dose of isoflavones, the changes in liver function, hematological parameters, and coagulation test were not different from those of the control subjects.<sup>(<xref rid="B38" ref-type="bibr">38</xref>)</sup> The dose of isoflavones administered in this clinical study corresponds to less than half of the estimated dose of daidzein that can exert anti-influenza activity. The duration of daidzein administration for the treatment of influenza infection is thought to be 5 days. Further research is necessary to examine the safety of short-term high dose administration of daidzein.</p><p>The inhibitory mechanism of daidzein against influenza replication does not involve the inhibition of viral enzymes, such as neuraminidase and cap-dependent endonuclease, which are encoded by the influenza virus genome, but rather involves enzyme activation in infected host cells. The results of the present study provide a foundation for developing anti-influenza treatments with a novel mode of action, as well as for developing preventive strategies against influenza infection using food ingredients.</p></sec><sec><title>Author Contributions</title><p>YH, RS, MS, NI, and RM performed experiments and analyzed the data; AO provided new tools and reagents; YI conceived and supervised the study; YI designed experiments and wrote the manuscript.</p></sec></body><back><ack><title>Acknowledgments</title><p>This work was supported by a Grant-in-Aid for Scientific Research (C) (grant numbers 24501017, 18K11117, and 19K11659) from the Japan Society for the Promotion of Science (KAKENHI).</p></ack><glossary><title>Abbreviations</title><def-list><def-item><term>7β-OHCh</term><def><p>7β-hydroxycholesterol</p></def></def-item><def-item><term>FFRA</term><def><p>focus-forming reduction assay</p></def></def-item><def-item><term>FFU</term><def><p>focus-forming unit</p></def></def-item><def-item><term>FLAP</term><def><p>5-lipoxygenase-activating protein</p></def></def-item><def-item><term>HETE</term><def><p>hydroxyeicosatetraenoic acid</p></def></def-item><def-item><term>HODE</term><def><p>hydroxyoctadecadienoic acid</p></def></def-item><def-item><term>HpETE</term><def><p>hydroperoxyeicosatetraenoic acid</p></def></def-item><def-item><term>8-iso-PGF2<sub>2α</sub></term><def><p>8-iso-prostaglandin F<sub>2α</sub></p></def></def-item><def-item><term>LC-MS/MS</term><def><p>liquid chromatography-tandem mass spectrometry</p></def></def-item><def-item><term>5-LOX</term><def><p>5-lipoxygenase</p></def></def-item><def-item><term>MDCK</term><def><p>Madin-Darby canine kidney</p></def></def-item><def-item><term>MOI</term><def><p>multiplicity of infection</p></def></def-item><def-item><term>PD1</term><def><p>protectin D1</p></def></def-item><def-item><term>PUFA</term><def><p>polyunsaturated fatty acid</p></def></def-item><def-item><term>ROS</term><def><p>reactive oxygen species</p></def></def-item><def-item><term>siRNA</term><def><p>short interfering RNA</p></def></def-item><def-item><term>tHODE</term><def><p>total hydroxyoctadecadienoic acid</p></def></def-item></def-list></glossary><sec sec-type="COI-statement"><title>Conflict of Interest</title><p>No potential conflicts of interest were disclosed.</p></sec><ref-list><title>References</title><ref id="B1"><label>1</label><mixed-citation publication-type="journal">Fiore AE, Uyeki TM, Broder K, et al.; Centers for Desease Control and Prevention (CDC). <article-title>Prevention and control of influenza with vaccines: recommendations of the Advisory Committee on Immunization Practices (ACIP), 2010</article-title>. <source>MMWR Recomm Rep</source><year>2010</year>; <volume>59</volume>: <fpage>1</fpage>–<lpage>62</lpage>.</mixed-citation></ref><ref id="B2"><label>2</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Belser</surname><given-names>JA</given-names></name>, <name name-style="western"><surname>Maines</surname><given-names>TR</given-names></name>, <name name-style="western"><surname>Tumpey</surname><given-names>TM</given-names></name></person-group><article-title>Importance of 1918 virus reconstruction to current assessments of pandemic risk</article-title>. <source>Virology</source><year>2018</year>; <volume>524</volume>: <fpage>45</fpage>–<lpage>55</lpage>.<pub-id pub-id-type="pmid">30142572</pub-id></mixed-citation></ref><ref id="B3"><label>3</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Chow</surname><given-names>EJ</given-names></name>, <name name-style="western"><surname>Doyle</surname><given-names>JD</given-names></name>, <name name-style="western"><surname>Uyeki</surname><given-names>TM</given-names></name></person-group><article-title>Influenza virus-related critical illness: prevention, diagnosis, treatment</article-title>. <source>Crit Care</source><year>2019</year>; <volume>23</volume>: <fpage>214</fpage>.<pub-id pub-id-type="pmid">31189475</pub-id></mixed-citation></ref><ref id="B4"><label>4</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Le</surname><given-names>QM</given-names></name>, <name name-style="western"><surname>Kiso</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Someya</surname><given-names>K</given-names></name>, <etal>et al</etal></person-group><article-title>Avian flu: isolation of drug-resistant H5N1 virus</article-title>. <source>Nature</source><year>2005</year>; <volume>437</volume>: <fpage>1108</fpage>.<pub-id pub-id-type="pmid">16228009</pub-id></mixed-citation></ref><ref id="B5"><label>5</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Bloom</surname><given-names>JD</given-names></name>, <name name-style="western"><surname>Gong</surname><given-names>LI</given-names></name>, <name name-style="western"><surname>Baltimore</surname><given-names>D</given-names></name></person-group><article-title>Permissive secondary mutations enable the evolution of influenza oseltamivir resistance</article-title>. <source>Science</source><year>2010</year>; <volume>328</volume>: <fpage>1272</fpage>–<lpage>1275</lpage>.<pub-id pub-id-type="pmid">20522774</pub-id></mixed-citation></ref><ref id="B6"><label>6</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Furuta</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Takahashi</surname><given-names>K</given-names></name>, <name name-style="western"><surname>Fukuda</surname><given-names>Y</given-names></name>, <etal>et al</etal></person-group><article-title><italic>In vitro</italic> and <italic>in vivo</italic> activities of anti-influenza virus compound T-705</article-title>. <source>Antimicrob Agents Chemother</source><year>2002</year>; <volume>46</volume>: <fpage>977</fpage>–<lpage>981</lpage>.<pub-id pub-id-type="pmid">11897578</pub-id></mixed-citation></ref><ref id="B7"><label>7</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Kiso</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Takahashi</surname><given-names>K</given-names></name>, <name name-style="western"><surname>Sakai-Tagawa</surname><given-names>Y</given-names></name>, <etal>et al</etal></person-group><article-title>T-705 (favipiravir) activity against lethal H5N1 influenza A viruses</article-title>. <source>Proc Natl Acad Sci U S A</source><year>2010</year>; <volume>107</volume>: <fpage>882</fpage>–<lpage>887</lpage>.<pub-id pub-id-type="pmid">20080770</pub-id></mixed-citation></ref><ref id="B8"><label>8</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Nagai</surname><given-names>E</given-names></name>, <name name-style="western"><surname>Iwai</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Koketsu</surname><given-names>R</given-names></name>, <etal>et al</etal></person-group><article-title>Inhibition of influenza virus replication by adlay tea</article-title>. <source>J Sci Food Agric</source><year>2018</year>; <volume>98</volume>: <fpage>1899</fpage>–<lpage>1905</lpage>.<pub-id pub-id-type="pmid">28902408</pub-id></mixed-citation></ref><ref id="B9"><label>9</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Christen</surname><given-names>S</given-names></name>, <name name-style="western"><surname>Peterhans</surname><given-names>E</given-names></name>, <name name-style="western"><surname>Stocker</surname><given-names>R</given-names></name></person-group><article-title>Antioxidant activities of some tryptophan metabolites: possible implication for inflammatory diseases</article-title>. <source>Proc Natl Acad Sci U S A</source><year>1990</year>; <volume>87</volume>: <fpage>2506</fpage>–<lpage>2510</lpage>.<pub-id pub-id-type="pmid">2320571</pub-id></mixed-citation></ref><ref id="B10"><label>10</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Jacoby</surname><given-names>DB</given-names></name>, <name name-style="western"><surname>Choi</surname><given-names>AM</given-names></name></person-group><article-title>Influenza virus induces expression of antioxidant genes in human epithelial cells</article-title>. <source>Free Radic Biol Med</source><year>1994</year>; <volume>16</volume>: <fpage>821</fpage>–<lpage>824</lpage>.<pub-id pub-id-type="pmid">8070686</pub-id></mixed-citation></ref><ref id="B11"><label>11</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Akaike</surname><given-names>T</given-names></name>, <name name-style="western"><surname>Noguchi</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Ijiri</surname><given-names>S</given-names></name>, <etal>et al</etal></person-group><article-title>Pathogenesis of influenza virus-induced pneumonia: involvement of both nitric oxide and oxygen radicals</article-title>. <source>Proc Natl Acad Sci U S A</source><year>1996</year>; <volume>93</volume>: <fpage>2448</fpage>–<lpage>2453</lpage>.<pub-id pub-id-type="pmid">8637894</pub-id></mixed-citation></ref><ref id="B12"><label>12</label><mixed-citation publication-type="journal">To EE, Erlich JR, Liong F, et al. Mitochondrial reactive oxygen species contribute to pathological inflammation during influenza A virus infection in mice. <source>Antioxid Redox Signal</source> 2019. DOI: 10.1089/ars.2019.7727.</mixed-citation></ref><ref id="B13"><label>13</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Jung</surname><given-names>KI</given-names></name>, <name name-style="western"><surname>Pyo</surname><given-names>CW</given-names></name>, <name name-style="western"><surname>Choi</surname><given-names>SY</given-names></name></person-group><article-title>Influenza A virus-induced autophagy contributes to enhancement of virus infectivity by SOD1 downregulation in alveolar epithelial cells</article-title>. <source>Biochem Biophys Res Commun</source><year>2018</year>; <volume>498</volume>: <fpage>960</fpage>–<lpage>966</lpage>.<pub-id pub-id-type="pmid">29548827</pub-id></mixed-citation></ref><ref id="B14"><label>14</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Franke</surname><given-names>AA</given-names></name>, <name name-style="western"><surname>Lai</surname><given-names>JF</given-names></name>, <name name-style="western"><surname>Halm</surname><given-names>BM</given-names></name></person-group><article-title>Absorption, distribution, metabolism, and excretion of isoflavonoids after soy intake</article-title>. <source>Arch Biochem Biophys</source><year>2014</year>; <volume>559</volume>: <fpage>24</fpage>–<lpage>28</lpage>.<pub-id pub-id-type="pmid">24946051</pub-id></mixed-citation></ref><ref id="B15"><label>15</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Sankar</surname><given-names>P</given-names></name>, <name name-style="western"><surname>Zachariah</surname><given-names>B</given-names></name>, <name name-style="western"><surname>Vickneshwaran</surname><given-names>V</given-names></name>, <name name-style="western"><surname>Jacob</surname><given-names>SE</given-names></name>, <name name-style="western"><surname>Sridhar</surname><given-names>MG</given-names></name></person-group><article-title> Amelioration of oxidative stress and insulin resistance by soy isoflavones (from Glycine max) in ovariectomized Wistar rats fed with high fat diet: the molecular mechanisms</article-title>. <source>Exp Gerontol</source><year>2015</year>; <volume>63</volume>: <fpage>67</fpage>–<lpage>75</lpage>.<pub-id pub-id-type="pmid">25660477</pub-id></mixed-citation></ref><ref id="B16"><label>16</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Chen</surname><given-names>W</given-names></name>, <name name-style="western"><surname>Ma</surname><given-names>X</given-names></name>, <name name-style="western"><surname>Lin</surname><given-names>Y</given-names></name>, <etal>et al</etal></person-group><article-title>Dietary supplementation with a high dose of daidzein enhances the antioxidant capacity in swine muscle but experts pro-oxidant function in liver and fat tissues</article-title>. <source>J Anim Sci Biotechnol</source><year>2016</year>; <volume>7</volume>: <fpage>43</fpage>.<pub-id pub-id-type="pmid">27486514</pub-id></mixed-citation></ref><ref id="B17"><label>17</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Jin</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Shen</surname><given-names>MH</given-names></name>, <name name-style="western"><surname>Jin</surname><given-names>MH</given-names></name>, <name name-style="western"><surname>Jin</surname><given-names>AH</given-names></name>, <name name-style="western"><surname>Yin</surname><given-names>XZ</given-names></name>, <name name-style="western"><surname>Quan</surname><given-names>JS</given-names></name></person-group><article-title>Hypoglycemic property of soy isoflavones from hypocotyl in Goto-Kakizaki diabetic rats</article-title>. <source>J Clin Biochem Nutr</source><year>2018</year>; <volume>62</volume>: <fpage>148</fpage>–<lpage>154</lpage>.<pub-id pub-id-type="pmid">29610554</pub-id></mixed-citation></ref><ref id="B18"><label>18</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Kuiper</surname><given-names>GG</given-names></name>, <name name-style="western"><surname>Carlsson</surname><given-names>B</given-names></name>, <name name-style="western"><surname>Grandien</surname><given-names>K</given-names></name>, <etal>et al</etal></person-group><article-title>Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta</article-title>. <source>Endocrinology</source><year>1997</year>; <volume>138</volume>: <fpage>863</fpage>–<lpage>870</lpage>.<pub-id pub-id-type="pmid">9048584</pub-id></mixed-citation></ref><ref id="B19"><label>19</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Kuiper</surname><given-names>GG</given-names></name>, <name name-style="western"><surname>Lemmen</surname><given-names>JG</given-names></name>, <name name-style="western"><surname>Carlsson</surname><given-names>B</given-names></name>, <etal>et al</etal></person-group><article-title>Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta</article-title>. <source>Endocrinology</source><year>1998</year>; <volume>139</volume>: <fpage>4252</fpage>–<lpage>4263</lpage>.<pub-id pub-id-type="pmid">9751507</pub-id></mixed-citation></ref><ref id="B20"><label>20</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Uifälean</surname><given-names>A</given-names></name>, <name name-style="western"><surname>Schneider</surname><given-names>S</given-names></name>, <name name-style="western"><surname>Ionescu</surname><given-names>C</given-names></name>, <name name-style="western"><surname>Lalk</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Iuga</surname><given-names>CA</given-names></name></person-group><article-title>Soy isoflavones and breast cancer cell lines: molecular mechanisms and future perspectives</article-title>. <source>Molecules</source><year>2015</year>; <volume>21</volume>: <fpage>E13</fpage>.<pub-id pub-id-type="pmid">26703550</pub-id></mixed-citation></ref><ref id="B21"><label>21</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Folman</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Pope</surname><given-names>GS</given-names></name></person-group><article-title>The interaction in the immature mouse of potent oestrogens with coumestrol, genistein and other utero-vaginotrophic compounds of low potency</article-title>. <source>J Endocrinol</source><year>1966</year>; <volume>34</volume>: <fpage>215</fpage>–<lpage>225</lpage>.<pub-id pub-id-type="pmid">5901836</pub-id></mixed-citation></ref><ref id="B22"><label>22</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Yan</surname><given-names>GR</given-names></name>, <name name-style="western"><surname>Xiao</surname><given-names>CL</given-names></name>, <name name-style="western"><surname>He</surname><given-names>GW</given-names></name>, <etal>et al</etal></person-group><article-title>Global phosphoproteomic effects of natural tyrosine kinase inhibitor, genistein, on signaling pathways</article-title>. <source>Proteomics</source><year>2010</year>; <volume>10</volume>: <fpage>976</fpage>–<lpage>986</lpage>.<pub-id pub-id-type="pmid">20049867</pub-id></mixed-citation></ref><ref id="B23"><label>23</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Hafez</surname><given-names>HS</given-names></name>, <name name-style="western"><surname>Ghareeb</surname><given-names>DA</given-names></name>, <name name-style="western"><surname>Saleh</surname><given-names>SR</given-names></name>, <etal>et al</etal></person-group><article-title>Neuroprotective effect of ipriflavone against scopolamine-induced memory impairment in rats</article-title>. <source>Psychopharmacology (Berl)</source><year>2017</year>; <volume>234</volume>: <fpage>3037</fpage>–<lpage>3053</lpage>.<pub-id pub-id-type="pmid">28733814</pub-id></mixed-citation></ref><ref id="B24"><label>24</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Kim</surname><given-names>MS</given-names></name>, <name name-style="western"><surname>Kim</surname><given-names>B</given-names></name>, <name name-style="western"><surname>Park</surname><given-names>H</given-names></name>, <etal>et al</etal></person-group><article-title>Long-term fermented soybean paste improves metabolic parameters associated with non-alcoholic fatty liver disease and insulin resistance in high-fat diet-induced obese mice</article-title>. <source>Biochem Biophys Res Commun</source><year>2018</year>; <volume>495</volume>: <fpage>1744</fpage>–<lpage>1751</lpage>.<pub-id pub-id-type="pmid">29222051</pub-id></mixed-citation></ref><ref id="B25"><label>25</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Yoshida</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Kodai</surname><given-names>S</given-names></name>, <name name-style="western"><surname>Takemura</surname><given-names>S</given-names></name>, <name name-style="western"><surname>Minamiyama</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Niki</surname><given-names>E</given-names></name></person-group><article-title>Simultaneous measurement of F2-isoprostane, hydroxyoctadecadienoic acid, hydroxyeicosatetraenoic acid, and hydroxycholesterols from physiological samples</article-title>. <source>Anal Biochem</source><year>2008</year>; <volume>379</volume>: <fpage>105</fpage>–<lpage>115</lpage>.<pub-id pub-id-type="pmid">18482573</pub-id></mixed-citation></ref><ref id="B26"><label>26</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Umeno</surname><given-names>A</given-names></name>, <name name-style="western"><surname>Shichiri</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Ishida</surname><given-names>N</given-names></name>, <etal>et al</etal></person-group><article-title>Singlet oxygen induced products of linoleates, 10- and 12-(Z,E)-hydroxyoctadecadienoic acids (HODE), can be potential biomarkers for early detection of type 2 diabetes</article-title>. <source>PLoS One</source><year>2013</year>; <volume>8</volume>: <fpage>e63542</fpage>.<pub-id pub-id-type="pmid">23691063</pub-id></mixed-citation></ref><ref id="B27"><label>27</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Shichiri</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Ishida</surname><given-names>N</given-names></name>, <name name-style="western"><surname>Hagihara</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Yoshida</surname><given-names>Y</given-names></name>, <name name-style="western"><surname>Kume</surname><given-names>A</given-names></name>, <name name-style="western"><surname>Suzuki</surname><given-names>H</given-names></name></person-group><article-title>Probucol induces the generation of lipid peroxidation products in erythrocytes and plasma of male cynomolgus macaques</article-title>. <source>J Clin Biochem Nutr</source><year>2019</year>; <volume>64</volume>: <fpage>129</fpage>–<lpage>142</lpage>.<pub-id pub-id-type="pmid">30936625</pub-id></mixed-citation></ref><ref id="B28"><label>28</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Lisovyy</surname><given-names>OO</given-names></name>, <name name-style="western"><surname>Dosenko</surname><given-names>VE</given-names></name>, <name name-style="western"><surname>Nagibin</surname><given-names>VS</given-names></name>, <etal>et al</etal></person-group><article-title>Cardioprotective effect of 5-lipoxygenase gene (ALOX5) silencing in ischemia-reperfusion</article-title>. <source>Acta Biochim Pol</source><year>2009</year>; <volume>56</volume>: <fpage>687</fpage>–<lpage>694</lpage>.<pub-id pub-id-type="pmid">20011686</pub-id></mixed-citation></ref><ref id="B29"><label>29</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Shichiri</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Kono</surname><given-names>N</given-names></name>, <name name-style="western"><surname>Shimanaka</surname><given-names>Y</given-names></name>, <etal>et al</etal></person-group><article-title>A novel role for α-tocopherol transfer protein (α-TTP) in protecting against chloroquine toxicity</article-title>. <source>J Biol Chem</source><year>2012</year>; <volume>287</volume>: <fpage>2926</fpage>–<lpage>2934</lpage>.<pub-id pub-id-type="pmid">22147702</pub-id></mixed-citation></ref><ref id="B30"><label>30</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Steinhilber</surname><given-names>D</given-names></name>, <name name-style="western"><surname>Fischer</surname><given-names>AS</given-names></name>, <name name-style="western"><surname>Metzner</surname><given-names>J</given-names></name>, <etal>et al</etal></person-group><article-title>5-lipoxygenase: underappreciated role of a pro-inflammatory enzyme in tumorigenesis</article-title>. <source>Front Pharmacol</source><year>2010</year>; <volume>1</volume>: <fpage>143</fpage>.<pub-id pub-id-type="pmid">21833182</pub-id></mixed-citation></ref><ref id="B31"><label>31</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Liu</surname><given-names>H</given-names></name>, <name name-style="western"><surname>Zhong</surname><given-names>H</given-names></name>, <name name-style="western"><surname>Leng</surname><given-names>L</given-names></name>, <name name-style="western"><surname>Jiang</surname><given-names>Z</given-names></name></person-group><article-title>Effects of soy isoflavone on hepatic steatosis in high fat-induced rats</article-title>. <source>J Clin Biochem Nutr</source><year>2017</year>; <volume>61</volume>: <fpage>85</fpage>–<lpage>90</lpage>.<pub-id pub-id-type="pmid">28955124</pub-id></mixed-citation></ref><ref id="B32"><label>32</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Bäck</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Yurdagul</surname><given-names>A</given-names><suffix>Jr</suffix></name>, <name name-style="western"><surname>Tabas</surname><given-names>I</given-names></name>, <name name-style="western"><surname>Öörni</surname><given-names>K</given-names></name>, <name name-style="western"><surname>Kovanen</surname><given-names>PT</given-names></name></person-group><article-title>Inflammation and its resolution in atherosclerosis: mediators and therapeutic opportunities</article-title>. <source>Nat Rev Cardiol</source><year>2019</year>; <volume>16</volume>: <fpage>389</fpage>–<lpage>406</lpage>.<pub-id pub-id-type="pmid">30846875</pub-id></mixed-citation></ref><ref id="B33"><label>33</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Serhan</surname><given-names>CN</given-names></name>, <name name-style="western"><surname>Dalli</surname><given-names>J</given-names></name>, <name name-style="western"><surname>Colas</surname><given-names>RA</given-names></name>, <name name-style="western"><surname>Winkler</surname><given-names>JW</given-names></name>, <name name-style="western"><surname>Chiang</surname><given-names>N</given-names></name></person-group><article-title>Protectins and maresins: new pro-resolving families of mediators in acute inflammation and resolution bioactive metabolome</article-title>. <source>Biochim Biophys Acta</source><year>2015</year>; <volume>1851</volume>: <fpage>397</fpage>–<lpage>413</lpage>.<pub-id pub-id-type="pmid">25139562</pub-id></mixed-citation></ref><ref id="B34"><label>34</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Morita</surname><given-names>M</given-names></name>, <name name-style="western"><surname>Kuba</surname><given-names>K</given-names></name>, <name name-style="western"><surname>Ichikawa</surname><given-names>A</given-names></name>, <etal>et al</etal></person-group><article-title>The lipid mediator protectin D1 inhibits influenza virus replication and improves severe influenza</article-title>. <source>Cell</source><year>2013</year>; <volume>153</volume>: <fpage>112</fpage>–<lpage>125</lpage>.<pub-id pub-id-type="pmid">23477864</pub-id></mixed-citation></ref><ref id="B35"><label>35</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Rådmark</surname><given-names>O</given-names></name>, <name name-style="western"><surname>Werz</surname><given-names>O</given-names></name>, <name name-style="western"><surname>Steinhilber</surname><given-names>D</given-names></name>, <name name-style="western"><surname>Samuelsson</surname><given-names>B</given-names></name></person-group><article-title>5-Lipoxygenase, a key enzyme for leukotriene biosynthesis in health and disease</article-title>. <source>Biochim Biophys Acta</source><year>2015</year>; <volume>1851</volume>: <fpage>331</fpage>–<lpage>339</lpage>.<pub-id pub-id-type="pmid">25152163</pub-id></mixed-citation></ref><ref id="B36"><label>36</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Recchiuti</surname><given-names>A</given-names></name>, <name name-style="western"><surname>Serhan</surname><given-names>CN</given-names></name></person-group><article-title>Pro-resolving ipid mediators (SPMs) and their actions in regulating miRNA in novel resolution circuits in inflammation</article-title>. <source>Front Immunol</source><year>2012</year>; <volume>3</volume>: <fpage>298</fpage>.<pub-id pub-id-type="pmid">23093949</pub-id></mixed-citation></ref><ref id="B37"><label>37</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>King</surname><given-names>RA</given-names></name>, <name name-style="western"><surname>Bursill</surname><given-names>DB</given-names></name></person-group><article-title>Plasma and urinary kinetics of the isoflavones daidzein and genistein after a single soy meal in humans</article-title>. <source>Am J Clin Nutr</source><year>1998</year>; <volume>67</volume>: <fpage>867</fpage>–<lpage>872</lpage>.<pub-id pub-id-type="pmid">9583843</pub-id></mixed-citation></ref><ref id="B38"><label>38</label><mixed-citation publication-type="journal"><person-group person-group-type="author"><name name-style="western"><surname>Cheng</surname><given-names>WC</given-names></name>, <name name-style="western"><surname>Lo</surname><given-names>SC</given-names></name>, <name name-style="western"><surname>Tsai</surname><given-names>KS</given-names></name>, <etal>et al</etal></person-group><article-title>Effects of high-dose phytoestrogens on circulating cellular microparticles and coagulation function in postmenopausal women</article-title>. <source>J Formos Med Assoc</source><year>2015</year>; <volume>114</volume>: <fpage>710</fpage>–<lpage>716</lpage>.<pub-id pub-id-type="pmid">24360978</pub-id></mixed-citation></ref></ref-list><sec sec-type="supplementary-material"><title>Supplementary Material</title><supplementary-material content-type="local-data" id="SF1"><caption><title>Supplemental Figure 1</title></caption><media mimetype="application" mime-subtype="pdf" xlink:href="jcbn19-70sf01.pdf" orientation="portrait" xlink:type="simple" id="d35e2795" position="anchor"></media></supplementary-material></sec></back><floats-group><fig id="F1" orientation="portrait" position="float"><label>Fig. 1</label><caption><p>Effect of daidzein on multiplication of influenza virus and on proliferation of MDCK cells. MDCK cells were inoculated with influenza A/PR/8/34 virus at a MOI of 0.001. (A) Concentration-dependent inhibitory effect of Daidzein on virus multiplication. Viral titers were determined at 24 h post-infection by focus-forming assays. (B) Cytotoxicity of daidzein. The cell viability of MDCK cells was determined at 24 h post-addition of daidzein by WST-8 assay. The dotted lines indicate a daidzein concentration of 275 µM. Data are presented as mean ± SD (<italic>n</italic> = 3). Data are representative of three independent experiments. <bold>*</bold><italic>p</italic><0.01; <bold>**</bold><italic>p</italic><0.05; <bold>***</bold><italic>p</italic><0.0025; <bold>****</bold><italic>p</italic><0.001.</p></caption><graphic xlink:href="jcbn19-70f01"></graphic></fig><fig id="F2" orientation="portrait" position="float"><label>Fig. 2</label><caption><p>Effect of daidzein on production of lipid peroxide. MDCK cells were inoculated with influenza A/PR/8/34 virus at a MOI of 0.001. Lipid fraction was harvested from MDCK cells at 24 h post-infection. (A) 7β-OHCh, (B) tHODE, (C) 8-iso-PGF<sub>2α</sub>, (D) 12-HETE, (E) 15-HETE, and (F) 5-HETE in cells. Data are presented as mean ± SD (<italic>n</italic> = 3). Data are representative of three independent experiments. <bold>*</bold><italic>p</italic><0.01, <bold>**</bold><italic>p</italic><0.005.</p></caption><graphic xlink:href="jcbn19-70f02"></graphic></fig><fig id="F3" orientation="portrait" position="float"><label>Fig. 3</label><caption><p>Effect of daidzein on lipoxygenase activity in MDCK cell lysate. The cell lysates were obtained from MDCK cells treated for 24 h with or without daidzein. AA was added to cell lysates to a final concentration at 120 µM and incubated for 180 min at 37°C with gentle mixing. After 180 min incubation, lipids were extracted and 5-, 12-, and 15-HETE and AA were measured by LC-MS/MS. The 5-, 12-, and 15-LOX activities in MDCK cell lysates were evaluated as the ratio of each HETE to AA. (A) 5-LOX activity was evaluated as 5-HETE/AA, (B) 12-LOX activity was evaluated as 12-HETE/AA, and (C) 15-LOX activity was evaluated as 15-HETE/AA. Data are presented as mean ± SD (<italic>n</italic> = 3). Data are representative of three independent experiments. <bold>*</bold><italic>p</italic><0.05.</p></caption><graphic xlink:href="jcbn19-70f03"></graphic></fig><fig id="F4" orientation="portrait" position="float"><label>Fig. 4</label><caption><p>Effect of 5-LOX inhibitor, zileuton, and 5-LOX siRNA transfection on multiplication of influenza virus and production of 5-HETE. MDCK cells were inoculated with influenza A/PR/8/34 virus at a MOI of 0.001. Viral titers were determined at 24 h post-infection by focus-forming assays. (A) Effect of zileuton on the titer of influenza virus. (B) Effect of zileuton on 5-HETE production. (C) Effect of 5-LOX siRNA on 5-LOX expression in whole cell lysate of MDCK cells treated with or without daidzein and with or without influenza virus infection. (D) Effect of 5-LOX siRNA on daidzein-induced inhibition of virus multiplication. Data are presented as mean ± SD. Data are representative of three independent experiments. <bold>*</bold><italic>p</italic><0.05, <bold>**</bold><italic>p</italic><0.025, <bold>***</bold><italic>p</italic><0.005, <bold>****</bold><italic>p</italic><0.001.</p></caption><graphic xlink:href="jcbn19-70f04"></graphic></fig><fig id="F5" orientation="portrait" position="float"><label>Fig. 5</label><caption><p>Effect of 5-HpETE on multiplication of influenza virus. MDCK cells were inoculated with influenza A/PR/8/34 virus at a MOI of 0.001. (A) Concentration-dependent inhibitory effect of 5-HpETE on virus multiplication. The inhibition rate of the virus titer was determined at 24 h post-infection by focus-forming assays. (B) Cytotoxicity of 5-HpETE. The survival rate of MDCK cells was determined at 24 h post-infection by WST-8 assay. Data are presented as mean ± SD (<italic>n</italic> = 3). Data are representative of three independent experiments.</p></caption><graphic xlink:href="jcbn19-70f05"></graphic></fig></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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