Wuhu Decoction Regulates Dendritic Cell Autophagy in the Treatment of Respiratory Syncytial Virus (RSV)-Induced Mouse Asthma by AMPK/ULK1 Signaling Pathway
Identifieur interne : 000693 ( Pmc/Curation ); précédent : 000692; suivant : 000694Wuhu Decoction Regulates Dendritic Cell Autophagy in the Treatment of Respiratory Syncytial Virus (RSV)-Induced Mouse Asthma by AMPK/ULK1 Signaling Pathway
Auteurs : Xingyu Chen [République populaire de Chine] ; Yinhe Luo [République populaire de Chine] ; Mengqing Wang [République populaire de Chine] ; Le Sun [République populaire de Chine] ; Kailing Huang [République populaire de Chine] ; Yan Li [République populaire de Chine] ; Yingying Chen [République populaire de Chine] ; Yi Ding [République populaire de Chine] ; Xin Zhang [République populaire de Chine] ; Luojia Jiao [République populaire de Chine] ; Jingyi Yang [République populaire de Chine] ; Ting Huang [République populaire de Chine]Source :
- Medical Science Monitor : International Medical Journal of Experimental and Clinical Research [ 1234-1010 ] ; 2019.
Abstract
Dendritic cell autophagy plays a pivotal role in asthma. Wuhu decoction can significantly improve respiratory syncytial virus (RSV) bronchiolitis and delay its development into asthma. The aim of the present study was to explore the therapeutic effect and mechanism of Wuhu decoction on RSV -induced asthma in mice.
Establishment of asthmatic mice model was induced by RSV. Hematoxylin-eosin staining, periodic acid-Schiff (PAS) staining, and Masson trichrome staining were performed to observe pathological changes in the lungs. The levels of CD4+ T, CD8+ T, and CD4+ CD25+ T in blood were analyzed by flow cytometry. The contents of interleukin (IL)-4, interferon-gamma (IFN-γ), IL-10, and IL-13 in serum were measured by enzyme-linked immunosorbent assay (ELISA). The number of autophagosomes in dendritic cells (DCs) of lung tissue was observed by transmission electron microscope. The DCs of lung tissue were isolated by magnetic bead sorting. The levels of LC3-II, Beclin-1, and LC3-I in DCs and MMP-9, TIMP-1, AMPK, p-AMPK, ULK1, and LK1 expression in lung tissues were detected by western blot. Real-time polymerase chain reaction (PCR) detected the expression of AMPK and ULK1 genes.
Wuhu decoction can effectively alleviate chronic airway inflammation and airway remodeling and reduce airway hyperresponsiveness. Moreover, Wuhu decoction can significantly enhance the level of autophagy in DCs of lung tissue and promote the expression of AMPK and ULK1 in lung tissue.
Wuhu decoction may improve the RSV-induced asthmatic symptoms by enhancing autophagy of DCs in lung tissue dependent on the AMPK/ULK1 signaling pathway.
Url:
DOI: 10.12659/MSM.917692
PubMed: 31325378
PubMed Central: 6662944
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Wuhu Decoction Regulates Dendritic Cell Autophagy in the Treatment of Respiratory Syncytial Virus (RSV)-Induced Mouse Asthma by AMPK/ULK1 Signaling Pathway</title>
<author><name sortKey="Chen, Xingyu" sort="Chen, Xingyu" uniqKey="Chen X" first="Xingyu" last="Chen">Xingyu Chen</name>
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<author><name sortKey="Luo, Yinhe" sort="Luo, Yinhe" uniqKey="Luo Y" first="Yinhe" last="Luo">Yinhe Luo</name>
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<author><name sortKey="Huang, Ting" sort="Huang, Ting" uniqKey="Huang T" first="Ting" last="Huang">Ting Huang</name>
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<country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country>
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<series><title level="j">Medical Science Monitor : International Medical Journal of Experimental and Clinical Research</title>
<idno type="ISSN">1234-1010</idno>
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>Dendritic cell autophagy plays a pivotal role in asthma. Wuhu decoction can significantly improve respiratory syncytial virus (RSV) bronchiolitis and delay its development into asthma. The aim of the present study was to explore the therapeutic effect and mechanism of Wuhu decoction on RSV -induced asthma in mice.</p>
</sec>
<sec><title>Material/Methods</title>
<p>Establishment of asthmatic mice model was induced by RSV. Hematoxylin-eosin staining, periodic acid-Schiff (PAS) staining, and Masson trichrome staining were performed to observe pathological changes in the lungs. The levels of CD4<sup>+</sup>
T, CD8<sup>+</sup>
T, and CD4<sup>+</sup>
CD25<sup>+</sup>
T in blood were analyzed by flow cytometry. The contents of interleukin (IL)-4, interferon-gamma (IFN-γ), IL-10, and IL-13 in serum were measured by enzyme-linked immunosorbent assay (ELISA). The number of autophagosomes in dendritic cells (DCs) of lung tissue was observed by transmission electron microscope. The DCs of lung tissue were isolated by magnetic bead sorting. The levels of LC3-II, Beclin-1, and LC3-I in DCs and MMP-9, TIMP-1, AMPK, p-AMPK, ULK1, and LK1 expression in lung tissues were detected by western blot. Real-time polymerase chain reaction (PCR) detected the expression of AMPK and ULK1 genes.</p>
</sec>
<sec><title>Results</title>
<p>Wuhu decoction can effectively alleviate chronic airway inflammation and airway remodeling and reduce airway hyperresponsiveness. Moreover, Wuhu decoction can significantly enhance the level of autophagy in DCs of lung tissue and promote the expression of AMPK and ULK1 in lung tissue.</p>
</sec>
<sec><title>Conclusions</title>
<p>Wuhu decoction may improve the RSV-induced asthmatic symptoms by enhancing autophagy of DCs in lung tissue dependent on the AMPK/ULK1 signaling pathway.</p>
</sec>
</div>
</front>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Med Sci Monit</journal-id>
<journal-id journal-id-type="iso-abbrev">Med. Sci. Monit</journal-id>
<journal-id journal-id-type="publisher-id">Medical Science Monitor</journal-id>
<journal-title-group><journal-title>Medical Science Monitor : International Medical Journal of Experimental and Clinical Research</journal-title>
</journal-title-group>
<issn pub-type="ppub">1234-1010</issn>
<issn pub-type="epub">1643-3750</issn>
<publisher><publisher-name>International Scientific Literature, Inc.</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">31325378</article-id>
<article-id pub-id-type="pmc">6662944</article-id>
<article-id pub-id-type="doi">10.12659/MSM.917692</article-id>
<article-id pub-id-type="publisher-id">917692</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Animal Study</subject>
</subj-group>
</article-categories>
<title-group><article-title>Wuhu Decoction Regulates Dendritic Cell Autophagy in the Treatment of Respiratory Syncytial Virus (RSV)-Induced Mouse Asthma by AMPK/ULK1 Signaling Pathway</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Chen</surname>
<given-names>Xingyu</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn1-medscimonit-25-5389">A</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><name><surname>Luo</surname>
<given-names>Yinhe</given-names>
</name>
<xref ref-type="aff" rid="af2-medscimonit-25-5389">2</xref>
<xref ref-type="author-notes" rid="fn1-medscimonit-25-5389">A</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><name><surname>Wang</surname>
<given-names>Mengqing</given-names>
</name>
<xref ref-type="aff" rid="af3-medscimonit-25-5389">3</xref>
<xref ref-type="author-notes" rid="fn2-medscimonit-25-5389">B</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Sun</surname>
<given-names>Le</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn4-medscimonit-25-5389">D</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Huang</surname>
<given-names>Kailing</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn3-medscimonit-25-5389">C</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Yan</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn4-medscimonit-25-5389">D</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chen</surname>
<given-names>Yingying</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn4-medscimonit-25-5389">D</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ding</surname>
<given-names>Yi</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn5-medscimonit-25-5389">E</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zhang</surname>
<given-names>Xin</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn5-medscimonit-25-5389">E</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Jiao</surname>
<given-names>Luojia</given-names>
</name>
<xref ref-type="aff" rid="af1-medscimonit-25-5389">1</xref>
<xref ref-type="author-notes" rid="fn5-medscimonit-25-5389">E</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yang</surname>
<given-names>Jingyi</given-names>
</name>
<xref ref-type="aff" rid="af3-medscimonit-25-5389">3</xref>
<xref ref-type="author-notes" rid="fn5-medscimonit-25-5389">E</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Huang</surname>
<given-names>Ting</given-names>
</name>
<xref ref-type="aff" rid="af3-medscimonit-25-5389">3</xref>
<xref ref-type="author-notes" rid="fn6-medscimonit-25-5389">F</xref>
</contrib>
</contrib-group>
<aff id="af1-medscimonit-25-5389"><label>1</label>
Hunan University of Chinese Medicine, Changsha, Hunan, P.R. China</aff>
<aff id="af2-medscimonit-25-5389"><label>2</label>
College of Integrated Traditional Chinese and Western Medicine of Hunan University of Chinese Medicine, Changsha, Hunan, P.R. China</aff>
<aff id="af3-medscimonit-25-5389"><label>3</label>
The Domestic First-Class Discipline Construction Project of Chinese Medicine of Hunan University of Chinese Medicine, Changsha, Hunan, P.R. China</aff>
<author-notes><corresp id="c1-medscimonit-25-5389">Corresponding Authors: Yinhe Luo, e-mail: <email>1286313109@qq.com</email>
, Mengqing Wang, e-mail: <email>wmqwmq2009@sina.com</email>
</corresp>
<fn id="fn1-medscimonit-25-5389"><label>A</label>
<p>Study Design</p>
</fn>
<fn id="fn2-medscimonit-25-5389"><label>B</label>
<p>Data Collection</p>
</fn>
<fn id="fn3-medscimonit-25-5389"><label>C</label>
<p>Statistical Analysis</p>
</fn>
<fn id="fn4-medscimonit-25-5389"><label>D</label>
<p>Data Interpretation</p>
</fn>
<fn id="fn5-medscimonit-25-5389"><label>E</label>
<p>Manuscript Preparation</p>
</fn>
<fn id="fn6-medscimonit-25-5389"><label>F</label>
<p>Literature Search</p>
</fn>
<fn id="fn7-medscimonit-25-5389"><label>G</label>
<p>Funds Collection</p>
</fn>
</author-notes>
<pub-date pub-type="collection"><year>2019</year>
</pub-date>
<pub-date pub-type="epub"><day>20</day>
<month>7</month>
<year>2019</year>
</pub-date>
<volume>25</volume>
<fpage>5389</fpage>
<lpage>5400</lpage>
<history><date date-type="received"><day>22</day>
<month>5</month>
<year>2019</year>
</date>
<date date-type="accepted"><day>01</day>
<month>7</month>
<year>2019</year>
</date>
</history>
<permissions><copyright-statement>© Med Sci Monit, 2019</copyright-statement>
<copyright-year>2019</copyright-year>
<license license-type="open-access"><license-p>This work is licensed under Creative Common Attribution-NonCommercial-NoDerivatives 4.0 International (<ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by-nc-nd/4.0/">CC BY-NC-ND 4.0</ext-link>
)</license-p>
</license>
</permissions>
<abstract><sec><title>Background</title>
<p>Dendritic cell autophagy plays a pivotal role in asthma. Wuhu decoction can significantly improve respiratory syncytial virus (RSV) bronchiolitis and delay its development into asthma. The aim of the present study was to explore the therapeutic effect and mechanism of Wuhu decoction on RSV -induced asthma in mice.</p>
</sec>
<sec><title>Material/Methods</title>
<p>Establishment of asthmatic mice model was induced by RSV. Hematoxylin-eosin staining, periodic acid-Schiff (PAS) staining, and Masson trichrome staining were performed to observe pathological changes in the lungs. The levels of CD4<sup>+</sup>
T, CD8<sup>+</sup>
T, and CD4<sup>+</sup>
CD25<sup>+</sup>
T in blood were analyzed by flow cytometry. The contents of interleukin (IL)-4, interferon-gamma (IFN-γ), IL-10, and IL-13 in serum were measured by enzyme-linked immunosorbent assay (ELISA). The number of autophagosomes in dendritic cells (DCs) of lung tissue was observed by transmission electron microscope. The DCs of lung tissue were isolated by magnetic bead sorting. The levels of LC3-II, Beclin-1, and LC3-I in DCs and MMP-9, TIMP-1, AMPK, p-AMPK, ULK1, and LK1 expression in lung tissues were detected by western blot. Real-time polymerase chain reaction (PCR) detected the expression of AMPK and ULK1 genes.</p>
</sec>
<sec><title>Results</title>
<p>Wuhu decoction can effectively alleviate chronic airway inflammation and airway remodeling and reduce airway hyperresponsiveness. Moreover, Wuhu decoction can significantly enhance the level of autophagy in DCs of lung tissue and promote the expression of AMPK and ULK1 in lung tissue.</p>
</sec>
<sec><title>Conclusions</title>
<p>Wuhu decoction may improve the RSV-induced asthmatic symptoms by enhancing autophagy of DCs in lung tissue dependent on the AMPK/ULK1 signaling pathway.</p>
</sec>
</abstract>
<kwd-group><title>MeSH Keywords</title>
<kwd>Asthma</kwd>
<kwd>Autophagy</kwd>
<kwd>Dendritic Cells</kwd>
<kwd>Respiratory Syncytial Viruses</kwd>
</kwd-group>
</article-meta>
</front>
<floats-group><fig id="f1-medscimonit-25-5389" position="float"><label>Figure 1</label>
<caption><p>Wuhu decoction improves airway hyperresponsiveness in a mouse model of RSV-induced asthma. (<bold>A</bold>
) Flow chart for establishing an asthma mouse model using OVA and RSV. (<bold>B</bold>
) Detection of airway reactivity and verification of successful modeling. (<bold>C</bold>
) Airway responsiveness of each group of mice challenged with different concentrations of acetylcholine was determined. * <italic>P</italic>
<0.05, ** <italic>P</italic>
<0.01, compared with the control group. <sup>#</sup>
<italic>P</italic>
<0.05, <sup>##</sup>
<italic>P</italic>
<0.01, compared with the asthma group. Mean ± standard deviation, n=10. OVA – with chicken egg albumin; RSV – respiratory syncytial virus.</p>
</caption>
<graphic xlink:href="medscimonit-25-5389-g001"></graphic>
</fig>
<fig id="f2-medscimonit-25-5389" position="float"><label>Figure 2</label>
<caption><p>Wuhu decoction improved airway inflammation in asthma mouse model induced by RSV. (<bold>A</bold>
) Representative images of HE staining sections of mouse lung tissue, and the nuclei were stained blue with hematoxylin. The cytoplasm is stained with eosin in varying shades of pink to deep red. (<bold>B, C</bold>
) The number of CD4<sup>+</sup>
T, CD8<sup>+</sup>
T and CD4<sup>+</sup>
CD25<sup>+</sup>
T in the blood of each group was detected by flow cytometry, and the CD4<sup>+</sup>
T/CD8<sup>+</sup>
T ratio was calculated. (<bold>D</bold>
) Serum levels of IL-4, IFN-γ, IL-10, and IL-13 were determined by ELISA. * <italic>P</italic>
<0.05, ** <italic>P</italic>
<0.01, compared with the control group. <sup>#</sup>
<italic>P</italic>
<0.05, <sup>##</sup>
<italic>P</italic>
<0.01, compared with the asthma group, mean ± standard deviation, n=10. RSV – respiratory syncytial virus; HE – hematoxylin-eosin; ELISA – enzyme-linked immunosorbent assay.</p>
</caption>
<graphic xlink:href="medscimonit-25-5389-g002"></graphic>
</fig>
<fig id="f3-medscimonit-25-5389" position="float"><label>Figure 3</label>
<caption><p>Wuhu decoction improved airway remodeling in asthma mouse model induced by RSV. (<bold>A</bold>
) Representative images of PAS and Masson staining sections of mouse lung tissue. The cytoplasm of PAS staining bronchial epithelial cells was red, and the nucleus was blue. The nuclei are stained black from Masson staining; muscle fibers are red; and collagen fibers are blue. (<bold>B</bold>
) The positive area of Masson staining was blue, PAS staining results showed that the positive areas were red, and the Image ProPlus 6.0 system was used for quantitative analysis to calculate the collagen area under the airway basement membrane and the index of airway mucus reserve. (<bold>C</bold>
) Western blot analysis of the expressions of AMPK and p-AMPK in lung tissues of mice in each group. * <italic>P</italic>
<0.05, ** <italic>P</italic>
<0.01, compared with the control group. <sup>#</sup>
<italic>P</italic>
<0.05, <sup>##</sup>
<italic>P</italic>
<0.01, compared with the asthma group. mean ± standard deviation, n=3. RSV – respiratory syncytial virus, PAS – periodic acid-Schiff.</p>
</caption>
<graphic xlink:href="medscimonit-25-5389-g003"></graphic>
</fig>
<fig id="f4-medscimonit-25-5389" position="float"><label>Figure 4</label>
<caption><p>The effect of Wuhu decoction on the autophagy level of DCs in the lung tissue of asthmatic mice induced by RSV. (<bold>A</bold>
) observation of autophagosomes in DCs of lung tissues of each group by transmission electron microscopy. (<bold>B</bold>
) Western blot was used to detect the expressions of LC3-I, LC3-II, and Beclin-1 in the lung DCs of mice in each group separated by magnetic beads, and the values of LC3-II/LC3-I were determined. * <italic>P</italic>
<0.05, ** <italic>P</italic>
<0.01, compared with the control group. <sup>#</sup>
<italic>P</italic>
<0.05, <sup>##</sup>
<italic>P</italic>
<0.01, compared with the asthma group. Mean ± standard deviation, n=3. DCs – dendritic cells; RSV – respiratory syncytial virus</p>
</caption>
<graphic xlink:href="medscimonit-25-5389-g004"></graphic>
</fig>
<fig id="f5-medscimonit-25-5389" position="float"><label>Figure 5</label>
<caption><p>Effect of Wuhu decoction on the AMPK/ULK1 pathway-related proteins in lung tissues of asthmatic mice induced by RSV. (<bold>A</bold>
) Western blot was used to detect the expressions of AMPK and p-AMPK in the lung tissues of mice in each group. (<bold>B</bold>
) Expression of ULK1 and p-ulk1 in lung tissues of mice in each group was detected by Western blot. (<bold>C</bold>
) Real-time PCR was performed to detect the expressions of AMPK and ULK1 genes in the lung tissues of mice. * <italic>P</italic>
<0.05, ** <italic>P</italic>
<0.01, compared with the control group. <sup>#</sup>
<italic>P</italic>
<0.05, <sup>##</sup>
<italic>P</italic>
<0.01, compared with the asthma group. Mean ± standard deviation, n=3. RSV – respiratory syncytial virus; PCR – polymerase chain reaction.</p>
</caption>
<graphic xlink:href="medscimonit-25-5389-g005"></graphic>
</fig>
</floats-group>
</pmc>
</record>
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