Bioassay-Guided Isolation of Nigracin, Responsible for the Tissue Repair Properties of Drypetes Klainei Stem Bark.
Identifieur interne : 000477 ( Main/Curation ); précédent : 000476; suivant : 000478Bioassay-Guided Isolation of Nigracin, Responsible for the Tissue Repair Properties of Drypetes Klainei Stem Bark.
Auteurs : Gianluca Sferrazza [Italie] ; Marco Corti [Italie] ; Federica Andreola [Italie] ; Daniela Giovannini [Italie] ; Giuseppe Nicotera [Italie] ; Manuela Zonfrillo [Italie] ; Massimo Serra [Italie] ; Sara Tengattini [Italie] ; Enrica Calleri [Italie] ; Gloria Brusotti [Italie] ; Pasquale Pierimarchi [Italie] ; Annalucia Serafino [Italie]Source :
- Frontiers in pharmacology [ 1663-9812 ] ; 2019.
Abstract
Drypetes klainei Pierre ex Pax is used in Cameroon by Baka people in the wound healing process and for the treatment of burns. In a previous paper we demonstrated the ability of both water (WE) and defatted methanol (DME) extracts to accelerate scratch wound closure in fibroblast cultures, thus validating the traditional use of D. klainey stem bark in the treatment of skin lesions. In this work we carried out a bioassay-guided fractionation of the most active DME, which exhibited in vitro efficacy in accelerating wound healing process, in order to isolate and identify the compound/s responsible for the assessed biological activity. HPLC was used for the metabolite profiling of DME and fractions (analytical) and for the isolation of the bioactive compound (semi-preparative). MS analyses and NMR spectroscopy were used for identifying the isolated compound. The abilities of treatments in accelerating wound healing were studied on murine fibroblasts in terms of cell viability and cell migration (scratch wound-healing assay). The results obtained allowed to unambiguously identify the isolated bioactive compound as nigracin, a known phenolic glycoside firstly isolated and characterized from bark and leaves of Populus nigra in 1967. However, this is the first time that nigracin is identified in the Drypetes genus and that a wound healing activity is demonstrated for this molecule. Specifically, we demonstrated that nigracin significantly stimulates fibroblast growth and improves cell motility and wound closure of fibroblast monolayer in a dose-dependent manner, without any toxicity at the concentrations tested, and is still active at very low doses. This makes the molecule particularly attractive as a possible candidate for developing new therapeutic options for wound care.
DOI: 10.3389/fphar.2019.01541
PubMed: 32038234
PubMed Central: PMC6989535
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first="Marco" last="Corti">Marco Corti</name><affiliation wicri:level="1"><nlm:affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Drug Sciences, University of Pavia, Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Andreola, Federica" sort="Andreola, Federica" uniqKey="Andreola F" first="Federica" last="Andreola">Federica Andreola</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Institute of Translational Pharmacology-National Research Council of Italy, Rome</wicri:regionArea></affiliation></author><author><name sortKey="Giovannini, Daniela" sort="Giovannini, Daniela" uniqKey="Giovannini D" first="Daniela" last="Giovannini">Daniela Giovannini</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Institute of Translational Pharmacology-National Research Council of Italy, Rome</wicri:regionArea></affiliation></author><author><name sortKey="Nicotera, Giuseppe" sort="Nicotera, Giuseppe" uniqKey="Nicotera G" first="Giuseppe" last="Nicotera">Giuseppe Nicotera</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Institute of Translational Pharmacology-National Research Council of Italy, Rome</wicri:regionArea></affiliation></author><author><name sortKey="Zonfrillo, Manuela" sort="Zonfrillo, Manuela" uniqKey="Zonfrillo M" first="Manuela" last="Zonfrillo">Manuela Zonfrillo</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Institute of Translational Pharmacology-National Research Council of Italy, Rome</wicri:regionArea></affiliation></author><author><name sortKey="Serra, Massimo" sort="Serra, Massimo" uniqKey="Serra M" first="Massimo" last="Serra">Massimo Serra</name><affiliation wicri:level="1"><nlm:affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Drug Sciences, University of Pavia, Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Tengattini, Sara" sort="Tengattini, Sara" uniqKey="Tengattini S" first="Sara" last="Tengattini">Sara Tengattini</name><affiliation wicri:level="1"><nlm:affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Drug Sciences, University of Pavia, Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Calleri, Enrica" sort="Calleri, Enrica" uniqKey="Calleri E" first="Enrica" last="Calleri">Enrica Calleri</name><affiliation wicri:level="1"><nlm:affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Drug Sciences, University of Pavia, Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Brusotti, Gloria" sort="Brusotti, Gloria" uniqKey="Brusotti G" first="Gloria" last="Brusotti">Gloria Brusotti</name><affiliation wicri:level="1"><nlm:affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Drug Sciences, University of Pavia, Pavia</wicri:regionArea></affiliation></author><author><name sortKey="Pierimarchi, Pasquale" sort="Pierimarchi, Pasquale" uniqKey="Pierimarchi P" first="Pasquale" last="Pierimarchi">Pasquale Pierimarchi</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Institute of Translational Pharmacology-National Research Council of Italy, Rome</wicri:regionArea></affiliation></author><author><name sortKey="Serafino, Annalucia" sort="Serafino, Annalucia" uniqKey="Serafino A" first="Annalucia" last="Serafino">Annalucia Serafino</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Institute of Translational Pharmacology-National Research Council of Italy, Rome</wicri:regionArea></affiliation></author></analytic><series><title level="j">Frontiers in pharmacology</title><idno type="ISSN">1663-9812</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><i>Drypetes klainei</i> Pierre ex Pax is used in Cameroon by Baka people in the wound healing process and for the treatment of burns. In a previous paper we demonstrated the ability of both water (WE) and defatted methanol (DME) extracts to accelerate scratch wound closure in fibroblast cultures, thus validating the traditional use of <i>D. klainey</i> stem bark in the treatment of skin lesions. In this work we carried out a bioassay-guided fractionation of the most active DME, which exhibited <i>in vitro</i> efficacy in accelerating wound healing process, in order to isolate and identify the compound/s responsible for the assessed biological activity. HPLC was used for the metabolite profiling of DME and fractions (analytical) and for the isolation of the bioactive compound (semi-preparative). MS analyses and NMR spectroscopy were used for identifying the isolated compound. The abilities of treatments in accelerating wound healing were studied on murine fibroblasts in terms of cell viability and cell migration (scratch wound-healing assay). The results obtained allowed to unambiguously identify the isolated bioactive compound as nigracin, a known phenolic glycoside firstly isolated and characterized from bark and leaves of <i>Populus nigra</i> in 1967. However, this is the first time that nigracin is identified in the <i>Drypetes</i> genus and that a wound healing activity is demonstrated for this molecule. Specifically, we demonstrated that nigracin significantly stimulates fibroblast growth and improves cell motility and wound closure of fibroblast monolayer in a dose-dependent manner, without any toxicity at the concentrations tested, and is still active at very low doses. This makes the molecule particularly attractive as a possible candidate for developing new therapeutic options for wound care.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">32038234</PMID><DateRevised><Year>2020</Year><Month>09</Month><Day>28</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">1663-9812</ISSN><JournalIssue CitedMedium="Print"><Volume>10</Volume><PubDate><Year>2019</Year></PubDate></JournalIssue><Title>Frontiers in pharmacology</Title><ISOAbbreviation>Front Pharmacol</ISOAbbreviation></Journal><ArticleTitle>Bioassay-Guided Isolation of Nigracin, Responsible for the Tissue Repair Properties of <i>Drypetes Klainei</i> Stem Bark.</ArticleTitle><Pagination><MedlinePgn>1541</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.3389/fphar.2019.01541</ELocationID><Abstract><AbstractText><i>Drypetes klainei</i> Pierre ex Pax is used in Cameroon by Baka people in the wound healing process and for the treatment of burns. In a previous paper we demonstrated the ability of both water (WE) and defatted methanol (DME) extracts to accelerate scratch wound closure in fibroblast cultures, thus validating the traditional use of <i>D. klainey</i> stem bark in the treatment of skin lesions. In this work we carried out a bioassay-guided fractionation of the most active DME, which exhibited <i>in vitro</i> efficacy in accelerating wound healing process, in order to isolate and identify the compound/s responsible for the assessed biological activity. HPLC was used for the metabolite profiling of DME and fractions (analytical) and for the isolation of the bioactive compound (semi-preparative). MS analyses and NMR spectroscopy were used for identifying the isolated compound. The abilities of treatments in accelerating wound healing were studied on murine fibroblasts in terms of cell viability and cell migration (scratch wound-healing assay). The results obtained allowed to unambiguously identify the isolated bioactive compound as nigracin, a known phenolic glycoside firstly isolated and characterized from bark and leaves of <i>Populus nigra</i> in 1967. However, this is the first time that nigracin is identified in the <i>Drypetes</i> genus and that a wound healing activity is demonstrated for this molecule. Specifically, we demonstrated that nigracin significantly stimulates fibroblast growth and improves cell motility and wound closure of fibroblast monolayer in a dose-dependent manner, without any toxicity at the concentrations tested, and is still active at very low doses. This makes the molecule particularly attractive as a possible candidate for developing new therapeutic options for wound care.</AbstractText><CopyrightInformation>Copyright © 2020 Sferrazza, Corti, Andreola, Giovannini, Nicotera, Zonfrillo, Serra, Tengattini, Calleri, Brusotti, Pierimarchi and Serafino.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Sferrazza</LastName><ForeName>Gianluca</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Corti</LastName><ForeName>Marco</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Andreola</LastName><ForeName>Federica</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Giovannini</LastName><ForeName>Daniela</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nicotera</LastName><ForeName>Giuseppe</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zonfrillo</LastName><ForeName>Manuela</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Serra</LastName><ForeName>Massimo</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tengattini</LastName><ForeName>Sara</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Calleri</LastName><ForeName>Enrica</ForeName><Initials>E</Initials><AffiliationInfo><Affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Brusotti</LastName><ForeName>Gloria</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Department of Drug Sciences, University of Pavia, Pavia, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Pierimarchi</LastName><ForeName>Pasquale</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Serafino</LastName><ForeName>Annalucia</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Institute of Translational Pharmacology-National Research Council of Italy, Rome, Italy.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>01</Month><Day>23</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Front Pharmacol</MedlineTA><NlmUniqueID>101548923</NlmUniqueID><ISSNLinking>1663-9812</ISSNLinking></MedlineJournalInfo><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Drypetes klainei Pierre ex Pax</Keyword><Keyword MajorTopicYN="N">bark extract</Keyword><Keyword MajorTopicYN="N">bioassay-guided isolation</Keyword><Keyword MajorTopicYN="N">nigracin</Keyword><Keyword MajorTopicYN="N">wound healing</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>05</Month><Day>03</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>11</Month><Day>27</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>2</Month><Day>11</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>2</Month><Day>11</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>2</Month><Day>11</Day><Hour>6</Hour><Minute>1</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId 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