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Article: Calycosin promotes angiogenesis involving estrogen receptor and mitogen-activated protein kinase (MAPK) signaling pathway in zebrafish and HUVEC
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TitleCalycosin promotes angiogenesis involving estrogen receptor and mitogen-activated protein kinase (MAPK) signaling pathway in zebrafish and HUVEC
 
AuthorsTang, JY2
Li, S2
Li, ZH2
Zhang, ZJ2
Hu, G2
Cheang, LCV2
Alex, D2
Hoi, MPM2
Kwan, YW4
Chan, SW3
Leung, GPH1
Lee, SMY2
 
Issue Date2010
 
PublisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
 
CitationPlos One, 2010, v. 5 n. 7 [How to Cite?]
DOI: http://dx.doi.org/10.1371/journal.pone.0011822
 
AbstractBackground: Angiogenesis plays an important role in a wide range of physiological processes, and many diseases are associated with the dysregulation of angiogenesis. Radix Astragali is a Chinese medicinal herb commonly used for treating cardiovascular disorders and has been shown to possess angiogenic effect in previous studies but its active constituent and underlying mechanism remain unclear. The present study investigates the angiogenic effects of calycosin, a major isoflavonoid isolated from Radix Astragali, in vitro and in vivo.Methodology: Tg(fli1:EGFP) and Tg(fli1:nEGFP) transgenic zebrafish embryos were treated with different concentrations of calycosin (10, 30, 100 μM) from 72 hpf to 96 hpf prior morphological observation and angiogenesis phenotypes assessment. Zebrafish embryos were exposed to calycosin (10, 100 μM) from 72 hpf to 78 hpf before gene-expression analysis. The effects of VEGFR tyrosine kinase inhibitor on calycosin-induced angiogenesis were studied using 72 hpf Tg(fli1:EGFP) and Tg(fli1:nEGFP) zebrafish embryos. The pro-angiogenic effects of calycosin were compared with raloxifene and tamoxifen in 72 hpf Tg(fli1:EGFP) zebrafish embryos. The binding affinities of calycosin to estrogen receptors (ERs) were evaluated by cell-free and cell-based estrogen receptor binding assays. Human umbilical vein endothelial cell cultures (HUVEC) were pretreated with different concentrations of calycosin (3, 10, 30, 100 mM) for 48 h then tested for cell viability and tube formation. The role of MAPK signaling in calycosin-induced angiogenesis was evaluated using western blotting. Conclusion: Calycosin was shown to induce angiogenesis in human umbilical vein endothelial cell cultures (HUVEC) in vitro and zebrafish embryos in vivo via the up-regulation of vascular endothelial growth factor (VEGF), VEGFR1 and VEGFR2 mRNA expression. It was demonstrated that calycosin acted similar to other selective estrogen receptor modulators (SERMs), such as raloxifene and tamoxifen, by displaying selective potency and affinity to estrogen receptors ERa and ERb. Our results further indicated that calycosin promotes angiogenesis via activation of MAPK with the involvement of ERK1/2 and ER. Together, this study revealed, for the first time, that calycosin acts as a selective estrogen receptor modulator (SERM) to promote angiogenesis, at least in part through VEGF-VEGFR2 and MAPK signaling pathways. © 2010 Tang et al.
 
ISSN1932-6203
2012 Impact Factor: 3.73
2012 SCImago Journal Rankings: 1.512
 
DOIhttp://dx.doi.org/10.1371/journal.pone.0011822
 
PubMed Central IDPMC2912279
 
ISI Accession Number IDWOS:000280520200007
Funding AgencyGrant Number
Science and Technology Development Fund of Macau SAR045/2007/A3
Research Committee, University of MacauRG085
UL017/09-Y1
Funding Information:

This study is supported by grant from the Science and Technology Development Fund of Macau SAR (Ref. No. 045/2007/A3) and Research Committee, University of Macau (Ref. No. RG085 and UL017/09-Y1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorTang, JY
 
dc.contributor.authorLi, S
 
dc.contributor.authorLi, ZH
 
dc.contributor.authorZhang, ZJ
 
dc.contributor.authorHu, G
 
dc.contributor.authorCheang, LCV
 
dc.contributor.authorAlex, D
 
dc.contributor.authorHoi, MPM
 
dc.contributor.authorKwan, YW
 
dc.contributor.authorChan, SW
 
dc.contributor.authorLeung, GPH
 
dc.contributor.authorLee, SMY
 
dc.date.accessioned2011-08-26T14:26:05Z
 
dc.date.available2011-08-26T14:26:05Z
 
dc.date.issued2010
 
dc.description.abstractBackground: Angiogenesis plays an important role in a wide range of physiological processes, and many diseases are associated with the dysregulation of angiogenesis. Radix Astragali is a Chinese medicinal herb commonly used for treating cardiovascular disorders and has been shown to possess angiogenic effect in previous studies but its active constituent and underlying mechanism remain unclear. The present study investigates the angiogenic effects of calycosin, a major isoflavonoid isolated from Radix Astragali, in vitro and in vivo.Methodology: Tg(fli1:EGFP) and Tg(fli1:nEGFP) transgenic zebrafish embryos were treated with different concentrations of calycosin (10, 30, 100 μM) from 72 hpf to 96 hpf prior morphological observation and angiogenesis phenotypes assessment. Zebrafish embryos were exposed to calycosin (10, 100 μM) from 72 hpf to 78 hpf before gene-expression analysis. The effects of VEGFR tyrosine kinase inhibitor on calycosin-induced angiogenesis were studied using 72 hpf Tg(fli1:EGFP) and Tg(fli1:nEGFP) zebrafish embryos. The pro-angiogenic effects of calycosin were compared with raloxifene and tamoxifen in 72 hpf Tg(fli1:EGFP) zebrafish embryos. The binding affinities of calycosin to estrogen receptors (ERs) were evaluated by cell-free and cell-based estrogen receptor binding assays. Human umbilical vein endothelial cell cultures (HUVEC) were pretreated with different concentrations of calycosin (3, 10, 30, 100 mM) for 48 h then tested for cell viability and tube formation. The role of MAPK signaling in calycosin-induced angiogenesis was evaluated using western blotting. Conclusion: Calycosin was shown to induce angiogenesis in human umbilical vein endothelial cell cultures (HUVEC) in vitro and zebrafish embryos in vivo via the up-regulation of vascular endothelial growth factor (VEGF), VEGFR1 and VEGFR2 mRNA expression. It was demonstrated that calycosin acted similar to other selective estrogen receptor modulators (SERMs), such as raloxifene and tamoxifen, by displaying selective potency and affinity to estrogen receptors ERa and ERb. Our results further indicated that calycosin promotes angiogenesis via activation of MAPK with the involvement of ERK1/2 and ER. Together, this study revealed, for the first time, that calycosin acts as a selective estrogen receptor modulator (SERM) to promote angiogenesis, at least in part through VEGF-VEGFR2 and MAPK signaling pathways. © 2010 Tang et al.
 
dc.description.naturepublished_or_final_version
 
dc.identifier.citationPlos One, 2010, v. 5 n. 7 [How to Cite?]
DOI: http://dx.doi.org/10.1371/journal.pone.0011822
 
dc.identifier.doihttp://dx.doi.org/10.1371/journal.pone.0011822
 
dc.identifier.epagee11822
 
dc.identifier.hkuros189205
 
dc.identifier.isiWOS:000280520200007
Funding AgencyGrant Number
Science and Technology Development Fund of Macau SAR045/2007/A3
Research Committee, University of MacauRG085
UL017/09-Y1
Funding Information:

This study is supported by grant from the Science and Technology Development Fund of Macau SAR (Ref. No. 045/2007/A3) and Research Committee, University of Macau (Ref. No. RG085 and UL017/09-Y1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

 
dc.identifier.issn1932-6203
2012 Impact Factor: 3.73
2012 SCImago Journal Rankings: 1.512
 
dc.identifier.issue7
 
dc.identifier.pmcidPMC2912279
 
dc.identifier.pmid20686605
 
dc.identifier.scopuseid_2-s2.0-77955591123
 
dc.identifier.spagee11822
 
dc.identifier.urihttp://hdl.handle.net/10722/137486
 
dc.identifier.volume5
 
dc.languageeng
 
dc.publisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
 
dc.publisher.placeUnited States
 
dc.relation.ispartofPLoS ONE
 
dc.relation.referencesReferences in Scopus
 
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License
 
dc.subject.meshEndothelial Cells - cytology - metabolism
 
dc.subject.meshIsoflavones - genetics - metabolism
 
dc.subject.meshMitogen-Activated Protein Kinases - genetics - metabolism
 
dc.subject.meshNeovascularization, Physiologic - genetics - physiology
 
dc.subject.meshReceptors, Estrogen - genetics - metabolism
 
dc.titleCalycosin promotes angiogenesis involving estrogen receptor and mitogen-activated protein kinase (MAPK) signaling pathway in zebrafish and HUVEC
 
dc.typeArticle
 
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<contributor.author>Li, S</contributor.author>
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<contributor.author>Zhang, ZJ</contributor.author>
<contributor.author>Hu, G</contributor.author>
<contributor.author>Cheang, LCV</contributor.author>
<contributor.author>Alex, D</contributor.author>
<contributor.author>Hoi, MPM</contributor.author>
<contributor.author>Kwan, YW</contributor.author>
<contributor.author>Chan, SW</contributor.author>
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<description.abstract>Background: Angiogenesis plays an important role in a wide range of physiological processes, and many diseases are associated with the dysregulation of angiogenesis. Radix Astragali is a Chinese medicinal herb commonly used for treating cardiovascular disorders and has been shown to possess angiogenic effect in previous studies but its active constituent and underlying mechanism remain unclear. The present study investigates the angiogenic effects of calycosin, a major isoflavonoid isolated from Radix Astragali, in vitro and in vivo.Methodology: Tg(fli1:EGFP) and Tg(fli1:nEGFP) transgenic zebrafish embryos were treated with different concentrations of calycosin (10, 30, 100 &#956;M) from 72 hpf to 96 hpf prior morphological observation and angiogenesis phenotypes assessment. Zebrafish embryos were exposed to calycosin (10, 100 &#956;M) from 72 hpf to 78 hpf before gene-expression analysis. The effects of VEGFR tyrosine kinase inhibitor on calycosin-induced angiogenesis were studied using 72 hpf Tg(fli1:EGFP) and Tg(fli1:nEGFP) zebrafish embryos. The pro-angiogenic effects of calycosin were compared with raloxifene and tamoxifen in 72 hpf Tg(fli1:EGFP) zebrafish embryos. The binding affinities of calycosin to estrogen receptors (ERs) were evaluated by cell-free and cell-based estrogen receptor binding assays. Human umbilical vein endothelial cell cultures (HUVEC) were pretreated with different concentrations of calycosin (3, 10, 30, 100 mM) for 48 h then tested for cell viability and tube formation. The role of MAPK signaling in calycosin-induced angiogenesis was evaluated using western blotting. Conclusion: Calycosin was shown to induce angiogenesis in human umbilical vein endothelial cell cultures (HUVEC) in vitro and zebrafish embryos in vivo via the up-regulation of vascular endothelial growth factor (VEGF), VEGFR1 and VEGFR2 mRNA expression. It was demonstrated that calycosin acted similar to other selective estrogen receptor modulators (SERMs), such as raloxifene and tamoxifen, by displaying selective potency and affinity to estrogen receptors ERa and ERb. Our results further indicated that calycosin promotes angiogenesis via activation of MAPK with the involvement of ERK1/2 and ER. Together, this study revealed, for the first time, that calycosin acts as a selective estrogen receptor modulator (SERM) to promote angiogenesis, at least in part through VEGF-VEGFR2 and MAPK signaling pathways. &#169; 2010 Tang et al.</description.abstract>
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Author Affiliations
  1. The University of Hong Kong
  2. University of Macau
  3. Hong Kong Polytechnic University
  4. Chinese University of Hong Kong