Ligustrazine ameliorates acute kidney injury through downregulation of NOD2-mediated inflammation
Identifieur interne : 000602 ( Pmc/Curation ); précédent : 000601; suivant : 000603Ligustrazine ameliorates acute kidney injury through downregulation of NOD2-mediated inflammation
Auteurs : Guosheng Jiang ; Rui Xin ; Wendan Yuan ; Lixia Zhang ; Xianghui Meng ; Wangnan Sun ; Huirong Han ; Yun Hou ; Lin Wang [République populaire de Chine] ; Pengchao DuSource :
- International Journal of Molecular Medicine [ 1107-3756 ] ; 2020.
Abstract
Ligustrazine has been used to alleviate clinical acute kidney injury (AKI); however, the underlying molecular mechanisms are poorly understood. In order to further elucidate the molecular mechanism underlying its occurrence, the role of nucleotide-binding oligomerization domain-containing 2 (NOD2) in AKI was investigated in the present study, and the results indicated that ligustrazine exerts an important protective effect against AKI
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DOI: 10.3892/ijmm.2020.4464
PubMed: 31985025
PubMed Central: 7015130
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Guosheng Jiang<affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation><affiliation><nlm:aff id="af2-ijmm-45-03-0731">Center for Reproductive Medicine, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, The Key Laboratory of Reproductive Endocrinology, Shandong University, Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, Shandong 250021</nlm:aff><wicri:noCountry code="subfield">Shandong 250021</wicri:noCountry></affiliation><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation><affiliation><nlm:aff id="af3-ijmm-45-03-0731">Microwave Treatment Department, Central Hospital of Zibo (GaoQing Branch Courts), Zibo, Shandong 256300</nlm:aff><wicri:noCountry code="subfield">Shandong 256300</wicri:noCountry></affiliation><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation><affiliation><nlm:aff id="af4-ijmm-45-03-0731">Department of Anesthesiology and Shandong Provincial Medicine and Health Key Laboratory of Clinical Anesthesia, Weifang Medical University, Weifang, Shandong 261053</nlm:aff><wicri:noCountry code="subfield">Shandong 261053</wicri:noCountry></affiliation><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation>Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Ligustrazine ameliorates acute kidney injury through downregulation of NOD2-mediated inflammation</title><author><name sortKey="Jiang, Guosheng" sort="Jiang, Guosheng" uniqKey="Jiang G" first="Guosheng" last="Jiang">Guosheng Jiang</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Xin, Rui" sort="Xin, Rui" uniqKey="Xin R" first="Rui" last="Xin">Rui Xin</name><affiliation><nlm:aff id="af2-ijmm-45-03-0731">Center for Reproductive Medicine, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, The Key Laboratory of Reproductive Endocrinology, Shandong University, Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, Shandong 250021</nlm:aff><wicri:noCountry code="subfield">Shandong 250021</wicri:noCountry></affiliation></author><author><name sortKey="Yuan, Wendan" sort="Yuan, Wendan" uniqKey="Yuan W" first="Wendan" last="Yuan">Wendan Yuan</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Zhang, Lixia" sort="Zhang, Lixia" uniqKey="Zhang L" first="Lixia" last="Zhang">Lixia Zhang</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Meng, Xianghui" sort="Meng, Xianghui" uniqKey="Meng X" first="Xianghui" last="Meng">Xianghui Meng</name><affiliation><nlm:aff id="af3-ijmm-45-03-0731">Microwave Treatment Department, Central Hospital of Zibo (GaoQing Branch Courts), Zibo, Shandong 256300</nlm:aff><wicri:noCountry code="subfield">Shandong 256300</wicri:noCountry></affiliation></author><author><name sortKey="Sun, Wangnan" sort="Sun, Wangnan" uniqKey="Sun W" first="Wangnan" last="Sun">Wangnan Sun</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Han, Huirong" sort="Han, Huirong" uniqKey="Han H" first="Huirong" last="Han">Huirong Han</name><affiliation><nlm:aff id="af4-ijmm-45-03-0731">Department of Anesthesiology and Shandong Provincial Medicine and Health Key Laboratory of Clinical Anesthesia, Weifang Medical University, Weifang, Shandong 261053</nlm:aff><wicri:noCountry code="subfield">Shandong 261053</wicri:noCountry></affiliation></author><author><name sortKey="Hou, Yun" sort="Hou, Yun" uniqKey="Hou Y" first="Yun" last="Hou">Yun Hou</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Wang, Lin" sort="Wang, Lin" uniqKey="Wang L" first="Lin" last="Wang">Lin Wang</name><affiliation wicri:level="1"><nlm:aff id="af5-ijmm-45-03-0731">Department of Gerontology, The Second Hospital of Shandong University, Jinan, Shandong 250000, P.R. China</nlm:aff><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>Department of Gerontology, The Second Hospital of Shandong University, Jinan, Shandong 250000</wicri:regionArea></affiliation></author><author><name sortKey="Du, Pengchao" sort="Du, Pengchao" uniqKey="Du P" first="Pengchao" last="Du">Pengchao Du</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">31985025</idno><idno type="pmc">7015130</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7015130</idno><idno type="RBID">PMC:7015130</idno><idno type="doi">10.3892/ijmm.2020.4464</idno><date when="2020">2020</date><idno type="wicri:Area/Pmc/Corpus">000602</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000602</idno><idno type="wicri:Area/Pmc/Curation">000602</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000602</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Ligustrazine ameliorates acute kidney injury through downregulation of NOD2-mediated inflammation</title><author><name sortKey="Jiang, Guosheng" sort="Jiang, Guosheng" uniqKey="Jiang G" first="Guosheng" last="Jiang">Guosheng Jiang</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Xin, Rui" sort="Xin, Rui" uniqKey="Xin R" first="Rui" last="Xin">Rui Xin</name><affiliation><nlm:aff id="af2-ijmm-45-03-0731">Center for Reproductive Medicine, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, The Key Laboratory of Reproductive Endocrinology, Shandong University, Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, Shandong 250021</nlm:aff><wicri:noCountry code="subfield">Shandong 250021</wicri:noCountry></affiliation></author><author><name sortKey="Yuan, Wendan" sort="Yuan, Wendan" uniqKey="Yuan W" first="Wendan" last="Yuan">Wendan Yuan</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Zhang, Lixia" sort="Zhang, Lixia" uniqKey="Zhang L" first="Lixia" last="Zhang">Lixia Zhang</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Meng, Xianghui" sort="Meng, Xianghui" uniqKey="Meng X" first="Xianghui" last="Meng">Xianghui Meng</name><affiliation><nlm:aff id="af3-ijmm-45-03-0731">Microwave Treatment Department, Central Hospital of Zibo (GaoQing Branch Courts), Zibo, Shandong 256300</nlm:aff><wicri:noCountry code="subfield">Shandong 256300</wicri:noCountry></affiliation></author><author><name sortKey="Sun, Wangnan" sort="Sun, Wangnan" uniqKey="Sun W" first="Wangnan" last="Sun">Wangnan Sun</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Han, Huirong" sort="Han, Huirong" uniqKey="Han H" first="Huirong" last="Han">Huirong Han</name><affiliation><nlm:aff id="af4-ijmm-45-03-0731">Department of Anesthesiology and Shandong Provincial Medicine and Health Key Laboratory of Clinical Anesthesia, Weifang Medical University, Weifang, Shandong 261053</nlm:aff><wicri:noCountry code="subfield">Shandong 261053</wicri:noCountry></affiliation></author><author><name sortKey="Hou, Yun" sort="Hou, Yun" uniqKey="Hou Y" first="Yun" last="Hou">Yun Hou</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author><author><name sortKey="Wang, Lin" sort="Wang, Lin" uniqKey="Wang L" first="Lin" last="Wang">Lin Wang</name><affiliation wicri:level="1"><nlm:aff id="af5-ijmm-45-03-0731">Department of Gerontology, The Second Hospital of Shandong University, Jinan, Shandong 250000, P.R. China</nlm:aff><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>Department of Gerontology, The Second Hospital of Shandong University, Jinan, Shandong 250000</wicri:regionArea></affiliation></author><author><name sortKey="Du, Pengchao" sort="Du, Pengchao" uniqKey="Du P" first="Pengchao" last="Du">Pengchao Du</name><affiliation><nlm:aff id="af1-ijmm-45-03-0731">School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</nlm:aff><wicri:noCountry code="subfield">Shandong 264003</wicri:noCountry></affiliation></author></analytic><series><title level="j">International Journal of Molecular Medicine</title><idno type="ISSN">1107-3756</idno><idno type="eISSN">1791-244X</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>Ligustrazine has been used to alleviate clinical acute kidney injury (AKI); however, the underlying molecular mechanisms are poorly understood. In order to further elucidate the molecular mechanism underlying its occurrence, the role of nucleotide-binding oligomerization domain-containing 2 (NOD2) in AKI was investigated in the present study, and the results indicated that ligustrazine exerts an important protective effect against AKI <italic>in vivo</italic> by inhibiting the upregulation of NOD2 expression and reducing apoptosis of kidney cells following ischemia/reperfusion injury in rat models. Furthermore, the inhibitory role of ligustrazine on the upregulation of NOD2 and apoptosis of kidney cells induced by CoCl<sub>2</sub> and oxygen and glucose deprivation followed by reoxygenation was investigated in <italic>in vitro</italic> experiments. The effect of ligustrazine on NOD2 downregulation was partially blocked by inhibiting autophagy. To the best of our knowledge, the results of the present study are the first to provide evidence that ligustrazine can inhibit NOD2-mediated inflammation to protect against renal injury, which may be in part attributed to the induction of autophagy. These findings may help design and develop new approaches and therapeutic strategies for AKI to prevent the deterioration of renal function.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Eltzschig, Hk" uniqKey="Eltzschig H">HK Eltzschig</name></author><author><name sortKey="Eckle, T" uniqKey="Eckle T">T Eckle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rabb, H" uniqKey="Rabb H">H Rabb</name></author><author><name sortKey="Griffin, Md" uniqKey="Griffin M">MD Griffin</name></author><author><name sortKey="Mckay, Db" uniqKey="Mckay D">DB McKay</name></author><author><name sortKey="Swaminathan, S" uniqKey="Swaminathan S">S Swaminathan</name></author><author><name sortKey="Pickkers, P" uniqKey="Pickkers P">P Pickkers</name></author><author><name sortKey="Rosner, Mh" uniqKey="Rosner M">MH Rosner</name></author><author><name sortKey="Kellum, Ja" uniqKey="Kellum J">JA 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Int J Mol Med</journal-id><journal-id journal-id-type="iso-abbrev">Int. J. Mol. Med</journal-id><journal-id journal-id-type="publisher-id">IJMM</journal-id><journal-title-group><journal-title>International Journal of Molecular Medicine</journal-title></journal-title-group><issn pub-type="ppub">1107-3756</issn><issn pub-type="epub">1791-244X</issn><publisher><publisher-name>D.A. Spandidos</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">31985025</article-id><article-id pub-id-type="pmc">7015130</article-id><article-id pub-id-type="doi">10.3892/ijmm.2020.4464</article-id><article-id pub-id-type="publisher-id">ijmm-45-03-0731</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group></article-categories><title-group><article-title>Ligustrazine ameliorates acute kidney injury through downregulation of NOD2-mediated inflammation</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Jiang</surname><given-names>Guosheng</given-names></name><xref ref-type="aff" rid="af1-ijmm-45-03-0731">1</xref><xref rid="fn1-ijmm-45-03-0731" ref-type="author-notes">*</xref></contrib><contrib contrib-type="author"><name><surname>Xin</surname><given-names>Rui</given-names></name><xref ref-type="aff" rid="af2-ijmm-45-03-0731">2</xref><xref rid="fn1-ijmm-45-03-0731" ref-type="author-notes">*</xref></contrib><contrib contrib-type="author"><name><surname>Yuan</surname><given-names>Wendan</given-names></name><xref ref-type="aff" rid="af1-ijmm-45-03-0731">1</xref><xref rid="fn1-ijmm-45-03-0731" ref-type="author-notes">*</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Lixia</given-names></name><xref ref-type="aff" rid="af1-ijmm-45-03-0731">1</xref></contrib><contrib contrib-type="author"><name><surname>Meng</surname><given-names>Xianghui</given-names></name><xref ref-type="aff" rid="af3-ijmm-45-03-0731">3</xref></contrib><contrib contrib-type="author"><name><surname>Sun</surname><given-names>Wangnan</given-names></name><xref ref-type="aff" rid="af1-ijmm-45-03-0731">1</xref></contrib><contrib contrib-type="author"><name><surname>Han</surname><given-names>Huirong</given-names></name><xref ref-type="aff" rid="af4-ijmm-45-03-0731">4</xref></contrib><contrib contrib-type="author"><name><surname>Hou</surname><given-names>Yun</given-names></name><xref ref-type="aff" rid="af1-ijmm-45-03-0731">1</xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Lin</given-names></name><xref ref-type="aff" rid="af5-ijmm-45-03-0731">5</xref></contrib><contrib contrib-type="author"><name><surname>Du</surname><given-names>Pengchao</given-names></name><xref ref-type="aff" rid="af1-ijmm-45-03-0731">1</xref><xref ref-type="corresp" rid="c1-ijmm-45-03-0731"></xref></contrib></contrib-group><aff id="af1-ijmm-45-03-0731"><label>1</label>School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264003</aff><aff id="af2-ijmm-45-03-0731"><label>2</label>Center for Reproductive Medicine, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, The Key Laboratory of Reproductive Endocrinology, Shandong University, Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, Shandong 250021</aff><aff id="af3-ijmm-45-03-0731"><label>3</label>Microwave Treatment Department, Central Hospital of Zibo (GaoQing Branch Courts), Zibo, Shandong 256300</aff><aff id="af4-ijmm-45-03-0731"><label>4</label>Department of Anesthesiology and Shandong Provincial Medicine and Health Key Laboratory of Clinical Anesthesia, Weifang Medical University, Weifang, Shandong 261053</aff><aff id="af5-ijmm-45-03-0731"><label>5</label>Department of Gerontology, The Second Hospital of Shandong University, Jinan, Shandong 250000, P.R. China</aff><author-notes><corresp id="c1-ijmm-45-03-0731">Correspondence to: Dr Pengchao Du, School of Basic Medical Sciences, Binzhou Medical University, 346 Guanhai Road, Yantai, Shandong 264003, P.R. China, E-mail: <email>252983491@qq.com</email></corresp><fn id="fn1-ijmm-45-03-0731" fn-type="equal"><label>*</label><p>Contributed equally</p></fn></author-notes><pub-date pub-type="ppub"><month>3</month><year>2020</year></pub-date><pub-date pub-type="epub"><day>10</day><month>1</month><year>2020</year></pub-date><pub-date pub-type="pmc-release"><day>10</day><month>1</month><year>2020</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>45</volume><issue>3</issue><fpage>731</fpage><lpage>742</lpage><history><date date-type="received"><day>16</day><month>7</month><year>2019</year></date><date date-type="accepted"><day>25</day><month>11</month><year>2019</year></date></history><permissions><copyright-statement>Copyright: © Jiang et al.</copyright-statement><copyright-year>2020</copyright-year><license license-type="open-access"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by-nc-nd/4.0/">Creative Commons Attribution-NonCommercial-NoDerivs License</ext-link>, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.</license-p></license></permissions><abstract><p>Ligustrazine has been used to alleviate clinical acute kidney injury (AKI); however, the underlying molecular mechanisms are poorly understood. In order to further elucidate the molecular mechanism underlying its occurrence, the role of nucleotide-binding oligomerization domain-containing 2 (NOD2) in AKI was investigated in the present study, and the results indicated that ligustrazine exerts an important protective effect against AKI <italic>in vivo</italic> by inhibiting the upregulation of NOD2 expression and reducing apoptosis of kidney cells following ischemia/reperfusion injury in rat models. Furthermore, the inhibitory role of ligustrazine on the upregulation of NOD2 and apoptosis of kidney cells induced by CoCl<sub>2</sub> and oxygen and glucose deprivation followed by reoxygenation was investigated in <italic>in vitro</italic> experiments. The effect of ligustrazine on NOD2 downregulation was partially blocked by inhibiting autophagy. To the best of our knowledge, the results of the present study are the first to provide evidence that ligustrazine can inhibit NOD2-mediated inflammation to protect against renal injury, which may be in part attributed to the induction of autophagy. These findings may help design and develop new approaches and therapeutic strategies for AKI to prevent the deterioration of renal function.</p></abstract><kwd-group><kwd>nucleotide-binding oligomerization domain-containing 2</kwd><kwd>autophagy</kwd><kwd>ligustrazine</kwd><kwd>inflammation</kwd><kwd>acute kidney injury</kwd></kwd-group></article-meta></front><floats-group><fig id="f1-ijmm-45-03-0731" orientation="portrait" position="float"><label>Figure 1</label><caption><p>Ligustrazine protects against AKI by suppressing tubular damage and inflammatory responses following I/R in a rat model. (A) Representative photomicrographs showing the morphological changes of kidneys from different groups of rats. Red arrows indicated morphological kidney injury with scattered single cell necrosis or desquamation of proximal tubular cells with intact basement membranes, loss of the brush border and tubule dilation. Bars, 200 <italic>µ</italic>m at lower magnification (×40). Bars, 20 <italic>µ</italic>m at higher magnification (×400). (B) Quantitative assessment of tubular damage from different groups of rats. (C) RT-qPCR analysis showing the levels of proinflammatory mediators (TNF-α, IL-6 and MCP-1) in different groups of rats. (D) ELISA was used to evaluate the levels of proinflammatory mediators in different groups of rats. (E) Representative western blots and summarized data showing the protein levels of CD68 in kidneys from different groups. (F) Representative photomicrographs of CD68 IHC staining in kidneys from different groups. Bars, 120 <italic>µ</italic>m (magnification, ×200). <sup>*</sup>P<0.05 vs. sham-operated rats (n=9), <sup>#</sup>P<0.05 vs. I/R rats (n=9). Ligu, ligustrazine; AKI, acute kidney injury; I/R, ischemia/reperfusion; RT-qPCR, reverse transcription-quantitative PCR; TNF, tumor necrosis factor; IL, interleukin; MCP, monocyte chemoattractant protein.</p></caption><graphic xlink:href="IJMM-45-03-0731-g00"></graphic></fig><fig id="f2-ijmm-45-03-0731" orientation="portrait" position="float"><label>Figure 2</label><caption><p>Ligustrazine inhibits the upregulation of NOD2 following I/R in a rat model. (A) Relative quantitation of mRNA levels of NOD2 by RT-qPCR analysis in kidneys from different groups. (B) Representative western blots and summarized data showing the protein levels of NOD2 in kidneys from different groups. (C) Representative photomicrographs of NOD2 IHC staining in the tubules and glomeruli of kidneys from different groups. Bars, 100 <italic>µ</italic>m (magnification, ×200). (D) Quantitative analysis of IHC staining showing increased expression of NOD2 in the tubules and glomeruli of rats after I/R, which was significantly decreased with ligustrazine administration. <sup>*</sup>P<0.05 vs. sham-operated rats (n=9), <sup>#</sup>P<0.05 vs. I/R rats (n=9). Ligu, ligustrazine; I/R, ischemia/reperfusion; NOD2, nucleotide-binding oligomerization domain-containing 2; IHC, immunohistochemistry.</p></caption><graphic xlink:href="IJMM-45-03-0731-g01"></graphic></fig><fig id="f3-ijmm-45-03-0731" orientation="portrait" position="float"><label>Figure 3</label><caption><p>Ligustrazine inhibits kidney apoptosis after I/R. (A) TUNEL assays were performed to assess renal cell death. Nuclei were visualized using DAPI staining. Quantitative analysis of TUNEL+ cells (numbers per high-power field). Bars, 75 <italic>µ</italic>m. (B) Representative photomicrographs of caspase 3/cleaved caspase 3 immunohistochemical staining in kidneys from different groups. Bars, 20 <italic>µ</italic>m (magnification, ×400). (C) Representative western blots and summa-rized data showing the levels of the ratio of caspase 3/cleaved caspase 3 in kidneys from different groups. <sup>*</sup>P<0.05 vs. sham-operated rats (n=9), <sup>#</sup>P<0.05 vs. I/R rats (n=9). Ligu, ligustrazine; I/R, ischemia/reperfusion.</p></caption><graphic xlink:href="IJMM-45-03-0731-g02"></graphic></fig><fig id="f4-ijmm-45-03-0731" orientation="portrait" position="float"><label>Figure 4</label><caption><p>Ligustrazine inhibits NOD2 expression response to different hypoxia models in rat proximal tubule epithelial cells (NRK-52E cells). (A) Representative western blots and summarized data showing the protein levels of NOD2 in response to CoCl<sub>2</sub> treatment at different concentrations. The treatment duration of CoCl<sub>2</sub> was 12 h. (B) Representative western blots and summarized data showing the protein levels of NOD2 in response to different recovery times following OGD. (C) Representative western blots and summarized data showing the protein levels of NOD2 in response to ligustrazine (30 and 50 <italic>µ</italic>M) after CoCl<sub>2</sub> treatment. (D) Representative western blots and summarized data showing the protein levels of NOD2 in response to ligustrazine (30 and 50 <italic>µ</italic>M) after OGD treatment followed by reoxygenation (24 h). <sup>*</sup>P<0.05 vs. control, <sup>#</sup>P<0.05 vs. CoCl<sub>2</sub> treatment group (100 and 250 <italic>µ</italic>M), <sup>*,#</sup>P<0.05 vs. ligustrazine treatment group (30 <italic>µ</italic>M), <sup>**</sup>P<0.05 vs. OGD treatment group. All the experiments were performed in triplicate. Ligu, ligustrazine; I/R, ischemia/reperfusion; NOD2, nucleotide-binding oligomerization domain containing 2; OGD, oxygen and glucose deprivation; reoxy, reoxygenation.</p></caption><graphic xlink:href="IJMM-45-03-0731-g03"></graphic></fig><fig id="f5-ijmm-45-03-0731" orientation="portrait" position="float"><label>Figure 5</label><caption><p>Ligustrazine-mediated NOD2 downregulation is blocked by inhibiting autophagy in NRK-52E cells <italic>in vitro</italic>. (A) Representative western blots and summarized data showing the relative ratio of LC3A/B-II/I in response to CoCl<sub>2</sub> treatment at different concentrations. The treatment duration of CoCl<sub>2</sub> was 12 h. (B) Representative western blots and summarized data showing the relative ratio of LC3A/B-II/I in response to ligustrazine (30 and 50 <italic>µ</italic>M) after CoCl<sub>2</sub> treatment. The duration of CoCl<sub>2</sub> treatment was 12 h and the duration of ligustrazine treatment was 24 h. (C) Representative western blots and summarized data showing the protein levels of NOD2 in response to ligustrazine (50 <italic>µ</italic>M) after CoCl<sub>2</sub> treatment when the autophagy was inhibited by CQ. (D) Representative western blots and summarized data showing the protein levels of NOD2 in response to ligustrazine after OGD treatment followed by reoxygenation (24 h) when the autophagy was inhibited by CQ. <sup>*</sup>P<0.05 vs. control, <sup>#</sup>P<0.05 vs. CoCl<sub>2</sub> treatment group, <sup>**</sup>P<0.05 vs. ligustrazine (50 <italic>µ</italic>M) after CoCl<sub>2</sub> treatment group, <sup>##</sup>P<0.05 vs. OGD treatment followed by reoxygenation (24 h) group. <sup>##,*</sup>P<0.05 vs. ligustrazine (50 <italic>µ</italic>M) after OGD treatment followed by reoxygenation (24 h) group. All the experiments were performed in triplicate. Ligu, ligustrazine; I/R, ischemia/reperfusion; reoxy, reoxygenation; NOD2, nucleotide-binding oligomerization domain-containing 2; OGD, oxygen and glucose deprivation; CQ, chloroquine.</p></caption><graphic xlink:href="IJMM-45-03-0731-g04"></graphic></fig><fig id="f6-ijmm-45-03-0731" orientation="portrait" position="float"><label>Figure 6</label><caption><p>Ligustrazine-mediated downregulation of inflammation and apoptosis is blocked by inhibiting autophagy in NRK-52E cells <italic>in vitro</italic>. (A) RT-qPCR analysis demonstrated that ligustrazine treatment suppressed the levels of proinflammatory mediators, including TNF-α, IL-6 and MCP-1, and this effect was attenuated when autophagy was inhibited by CQ. The duration of reoxygenation was 24 h. (B) CCK-8 assays demonstrated that cell viability in response to ligustrazine after OGD followed by reoxygenation for 24 h was blocked when the autophagy was inhibited by CQ. (C) TUNEL assays showing the changes of cell death in response to ligustrazine after CoCl<sub>2</sub> treatment when autophagy was inhibited by CQ. The treatment duration of CoCl<sub>2</sub> was 12 h, and that of ligustrazine and CQ was 24 h. <sup>*</sup>P<0.05 vs. control, <sup>#</sup>P<0.05 vs. OGD treatment, <sup>**</sup>P<0.05 vs. ligustrazine treatment group. All the experiments were performed in triplicate. Ligu, ligustrazine; I/R, ischemia/reperfusion; reoxy, reoxygenation; OGD, oxygen and glucose deprivation; CQ, chloroquine; TNF, tumor necrosis factor; IL, interleukin; MCP, monocyte chemoattractant protein.</p></caption><graphic xlink:href="IJMM-45-03-0731-g05"></graphic></fig><table-wrap id="tI-ijmm-45-03-0731" orientation="portrait" position="float"><label>Table I</label><caption><p>Physical and biochemical parameters of experimental animals.</p></caption><table frame="hsides" rules="groups"><thead><tr><th valign="top" align="left" rowspan="1" colspan="1">Variables</th><th valign="top" align="center" rowspan="1" colspan="1">Sham</th><th valign="top" align="center" rowspan="1" colspan="1">I/R</th><th valign="top" align="center" rowspan="1" colspan="1">I/R + Ligu</th></tr></thead><tbody><tr><td valign="top" align="left" rowspan="1" colspan="1">Body weight (g)</td><td valign="top" align="center" rowspan="1" colspan="1">222±18</td><td valign="top" align="center" rowspan="1" colspan="1">220±20</td><td valign="top" align="center" rowspan="1" colspan="1">223±15</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1">Blood urea nitrogen (mmol/l)</td><td valign="top" align="center" rowspan="1" colspan="1">8.9±0.93</td><td valign="top" align="center" rowspan="1" colspan="1">42.1±4. 57<xref rid="tfn2-ijmm-45-03-0731" ref-type="table-fn">a</xref></td><td valign="top" align="center" rowspan="1" colspan="1">22.7±0.85<xref rid="tfn3-ijmm-45-03-0731" ref-type="table-fn">b</xref></td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1">Serum creatinine (<italic>µ</italic>mol/l)</td><td valign="top" align="center" rowspan="1" colspan="1">69.8±10.26</td><td valign="top" align="center" rowspan="1" colspan="1">206.1±25.73<xref rid="tfn2-ijmm-45-03-0731" ref-type="table-fn">a</xref></td><td valign="top" align="center" rowspan="1" colspan="1">145.6±15.49<xref rid="tfn3-ijmm-45-03-0731" ref-type="table-fn">b</xref></td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1">N</td><td valign="top" align="center" rowspan="1" colspan="1">9</td><td valign="top" align="center" rowspan="1" colspan="1">9</td><td valign="top" align="center" rowspan="1" colspan="1">9</td></tr></tbody></table><table-wrap-foot><fn id="tfn1-ijmm-45-03-0731"><p>Values are expressed as mean ± standard error of the mean.</p></fn><fn id="tfn2-ijmm-45-03-0731"><label>a</label><p>P<0.05 vs. scramble rats, was considered statistically significant.</p></fn><fn id="tfn3-ijmm-45-03-0731"><label>b</label><p>P<0.05 vs. I/R rats, was considered statistically significant. Sham, scramble rats; I/R, renal ischemia/reperfusion rats; I/R + Ligu, renal ischemia/reperfusion rats receiving ligustrazine treatment.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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