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Formononetin promotes angiogenesis through the estrogen receptor alpha-enhanced ROCK pathway.

Li S, Dang Y, Zhou X, Huang B, Huang X, Zhang Z, Kwan YW, Chan SW, Leung GP, Lee SM, Hoi MP - Sci Rep (2015)

Bottom Line: In addition, results from co-immunoprecipitation suggested formononetin induced cell migration via recruiting of ERα/ROCK-II activated complex formation.More interestingly, in zebrafish embryo we observed that formononetin significantly promoted angiogenic sproutings in the subintestinal vessels (SIVs) that could be completely abolished by ROCK inhibitor.In this study, we elucidated the underlying mechanisms that formononetin produced proangiogenesis effects through an ERα-enhanced ROCK-II signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine and Institute of Chinese Medical Sciences, University of Macau, Macao, China.

ABSTRACT
Formononetin is an isoflavone that has been shown to display estrogenic properties and induce angiogenesis activities. However, the interrelationship between the estrogenic properties and angiogenesis activities of formononetin are not well defined. In the present study, docking and enzymatic assay demonstrated that formononetin displayed direct binding to the ligand-binding domain (LBD) of estrogen receptor alpha (ERα) with an agonistic property. Results from Human Umbilical Vein Endothelial Cells (HUVEC) by using real-time migration xCELLigence system, immunofluorescence and western blotting provided strong evidences of formononetin induced endothelial cell migration and dramatic actin cytoskeleton spatial modification through ERα-enhanced-ROCK-II/MMP2/9 signaling pathways. In addition, results from co-immunoprecipitation suggested formononetin induced cell migration via recruiting of ERα/ROCK-II activated complex formation. More interestingly, in zebrafish embryo we observed that formononetin significantly promoted angiogenic sproutings in the subintestinal vessels (SIVs) that could be completely abolished by ROCK inhibitor. In this study, we elucidated the underlying mechanisms that formononetin produced proangiogenesis effects through an ERα-enhanced ROCK-II signaling pathways. Results from the present study also expand our knowledge about the enigmatic underlying mechanisms of phytoestrogenic compounds in the promotion of angiogenesis in relation to ERα and ROCK interaction in endothelial cells and their relationship with actin assembly and cell migration.

No MeSH data available.


Related in: MedlinePlus

The effects of ROCK inhibitor in formononetin-induced angiogenesis in zebrafish embryos in vivo.Tg (fli1: EGFP)y1 zebrafish embryos (48 hpf) were treated with (A) 0.1% DMSO (negative control), (B) formononetin (25 μM), (C) formononetin (50 μM), (D) H1152 (10 μM, ROCK inhibitor) alone, (E) co-treatment of H1152 (10 μM) +formononetin (25 μM), (F) co-treatment of H1152 (10 μM) +formononetin (50 μM). All treatments were for 24 h. (G) Quantitative analysis indicated the length of sprouting in SIVs for each group. Data are plotted as means ± SD, from three individual experiments. *p < 0.05, ***p < 0.001 vs. control group.
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f12: The effects of ROCK inhibitor in formononetin-induced angiogenesis in zebrafish embryos in vivo.Tg (fli1: EGFP)y1 zebrafish embryos (48 hpf) were treated with (A) 0.1% DMSO (negative control), (B) formononetin (25 μM), (C) formononetin (50 μM), (D) H1152 (10 μM, ROCK inhibitor) alone, (E) co-treatment of H1152 (10 μM) +formononetin (25 μM), (F) co-treatment of H1152 (10 μM) +formononetin (50 μM). All treatments were for 24 h. (G) Quantitative analysis indicated the length of sprouting in SIVs for each group. Data are plotted as means ± SD, from three individual experiments. *p < 0.05, ***p < 0.001 vs. control group.

Mentions: We further investigated the role of the ROCK pathway in formononetin-induced sproutings in vivo in zebrafish by using the ROCK inhibitior H1152. In agreement with the results in HUVECs, H1152 abolished the sproutings induced by formononetin significantly (Fig. 12D–F). These results suggested that formononetin induced angiogenesis in zebrafish embryos via multiple pathways and various steps in angiogenesis (e.g. cell proliferation and migration).


Formononetin promotes angiogenesis through the estrogen receptor alpha-enhanced ROCK pathway.

Li S, Dang Y, Zhou X, Huang B, Huang X, Zhang Z, Kwan YW, Chan SW, Leung GP, Lee SM, Hoi MP - Sci Rep (2015)

The effects of ROCK inhibitor in formononetin-induced angiogenesis in zebrafish embryos in vivo.Tg (fli1: EGFP)y1 zebrafish embryos (48 hpf) were treated with (A) 0.1% DMSO (negative control), (B) formononetin (25 μM), (C) formononetin (50 μM), (D) H1152 (10 μM, ROCK inhibitor) alone, (E) co-treatment of H1152 (10 μM) +formononetin (25 μM), (F) co-treatment of H1152 (10 μM) +formononetin (50 μM). All treatments were for 24 h. (G) Quantitative analysis indicated the length of sprouting in SIVs for each group. Data are plotted as means ± SD, from three individual experiments. *p < 0.05, ***p < 0.001 vs. control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645220&req=5

f12: The effects of ROCK inhibitor in formononetin-induced angiogenesis in zebrafish embryos in vivo.Tg (fli1: EGFP)y1 zebrafish embryos (48 hpf) were treated with (A) 0.1% DMSO (negative control), (B) formononetin (25 μM), (C) formononetin (50 μM), (D) H1152 (10 μM, ROCK inhibitor) alone, (E) co-treatment of H1152 (10 μM) +formononetin (25 μM), (F) co-treatment of H1152 (10 μM) +formononetin (50 μM). All treatments were for 24 h. (G) Quantitative analysis indicated the length of sprouting in SIVs for each group. Data are plotted as means ± SD, from three individual experiments. *p < 0.05, ***p < 0.001 vs. control group.
Mentions: We further investigated the role of the ROCK pathway in formononetin-induced sproutings in vivo in zebrafish by using the ROCK inhibitior H1152. In agreement with the results in HUVECs, H1152 abolished the sproutings induced by formononetin significantly (Fig. 12D–F). These results suggested that formononetin induced angiogenesis in zebrafish embryos via multiple pathways and various steps in angiogenesis (e.g. cell proliferation and migration).

Bottom Line: In addition, results from co-immunoprecipitation suggested formononetin induced cell migration via recruiting of ERα/ROCK-II activated complex formation.More interestingly, in zebrafish embryo we observed that formononetin significantly promoted angiogenic sproutings in the subintestinal vessels (SIVs) that could be completely abolished by ROCK inhibitor.In this study, we elucidated the underlying mechanisms that formononetin produced proangiogenesis effects through an ERα-enhanced ROCK-II signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine and Institute of Chinese Medical Sciences, University of Macau, Macao, China.

ABSTRACT
Formononetin is an isoflavone that has been shown to display estrogenic properties and induce angiogenesis activities. However, the interrelationship between the estrogenic properties and angiogenesis activities of formononetin are not well defined. In the present study, docking and enzymatic assay demonstrated that formononetin displayed direct binding to the ligand-binding domain (LBD) of estrogen receptor alpha (ERα) with an agonistic property. Results from Human Umbilical Vein Endothelial Cells (HUVEC) by using real-time migration xCELLigence system, immunofluorescence and western blotting provided strong evidences of formononetin induced endothelial cell migration and dramatic actin cytoskeleton spatial modification through ERα-enhanced-ROCK-II/MMP2/9 signaling pathways. In addition, results from co-immunoprecipitation suggested formononetin induced cell migration via recruiting of ERα/ROCK-II activated complex formation. More interestingly, in zebrafish embryo we observed that formononetin significantly promoted angiogenic sproutings in the subintestinal vessels (SIVs) that could be completely abolished by ROCK inhibitor. In this study, we elucidated the underlying mechanisms that formononetin produced proangiogenesis effects through an ERα-enhanced ROCK-II signaling pathways. Results from the present study also expand our knowledge about the enigmatic underlying mechanisms of phytoestrogenic compounds in the promotion of angiogenesis in relation to ERα and ROCK interaction in endothelial cells and their relationship with actin assembly and cell migration.

No MeSH data available.


Related in: MedlinePlus