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Beta-Adrenoceptor Activation Reduces Both Dermal Microvascular Endothelial Cell Migration via a cAMP-Dependent Mechanism and Wound Angiogenesis.

O'Leary AP, Fox JM, Pullar CE - J. Cell. Physiol. (2015)

Bottom Line: They play a role in wound repair but their specific role in angiogenesis is unknown.In this study, a range of in vitro assays (single cell migration, scratch wound healing, ELISAs for angiogenic growth factors and tubule formation) were performed with human dermal microvascular endothelial cells (HDMEC) to investigate and dissect mechanisms underpinning β-AR-mediated modulation of angiogenesis in chick chorioallantoic membranes (CAM) and murine excisional skin wounds. β-AR activation reduced HDMEC migration via cyclic adenosine monophosphate (cAMP)-dependent and protein kinase A (PKA)-independent mechanisms as demonstrated through use of an EPAC agonist that auto-inhibited the cAMP-mediated β-AR transduced reduction in HDMEC motility; a PKA inhibitor was, conversely, ineffective.In more complex environments, β-AR activation delayed HDMEC tubule formation and decreased angiogenesis both in the CAM assay and in murine excisional skin wounds in vivo. β-AR activation reduced HDMEC function in vitro and angiogenesis in vivo; therefore, β-AR agonists could be promising anti-angiogenic modulators in skin.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

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β-AR activation delays HDMEC scratch wound closure. Scratch wound assays were performed as described in the methods. Cells were treated with media alone or media containing 10 µM Iso. Demarcated areas of each well were photographed 0, 6, 12, 24 and 32 h later. Images representative of control and Iso-treated wounds at time 0 and 24 h are presented; scale bar = 200 μm (A). Combined data from 5–6 independent experiments using 3 s separate cell strains is shown after averaging, statistical analysis and graphical representation with the bars representing the means ± SEM (control N = 6; Iso N = 5) (**P < 0.01) (B).
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fig03: β-AR activation delays HDMEC scratch wound closure. Scratch wound assays were performed as described in the methods. Cells were treated with media alone or media containing 10 µM Iso. Demarcated areas of each well were photographed 0, 6, 12, 24 and 32 h later. Images representative of control and Iso-treated wounds at time 0 and 24 h are presented; scale bar = 200 μm (A). Combined data from 5–6 independent experiments using 3 s separate cell strains is shown after averaging, statistical analysis and graphical representation with the bars representing the means ± SEM (control N = 6; Iso N = 5) (**P < 0.01) (B).

Mentions: To investigate wound angiogenesis in vitro, the scratch wound assay was used to determine the effect of Iso on HDMEC migration from a monolayer wound edge. β-AR activation delayed wound closure at 24 h (Fig. 3A/B). After 32 h, control wounds were 92% healed, while closure of β-AR agonist-treated wounds were significantly delayed by 25% (Fig. 3B).


Beta-Adrenoceptor Activation Reduces Both Dermal Microvascular Endothelial Cell Migration via a cAMP-Dependent Mechanism and Wound Angiogenesis.

O'Leary AP, Fox JM, Pullar CE - J. Cell. Physiol. (2015)

β-AR activation delays HDMEC scratch wound closure. Scratch wound assays were performed as described in the methods. Cells were treated with media alone or media containing 10 µM Iso. Demarcated areas of each well were photographed 0, 6, 12, 24 and 32 h later. Images representative of control and Iso-treated wounds at time 0 and 24 h are presented; scale bar = 200 μm (A). Combined data from 5–6 independent experiments using 3 s separate cell strains is shown after averaging, statistical analysis and graphical representation with the bars representing the means ± SEM (control N = 6; Iso N = 5) (**P < 0.01) (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263239&req=5

fig03: β-AR activation delays HDMEC scratch wound closure. Scratch wound assays were performed as described in the methods. Cells were treated with media alone or media containing 10 µM Iso. Demarcated areas of each well were photographed 0, 6, 12, 24 and 32 h later. Images representative of control and Iso-treated wounds at time 0 and 24 h are presented; scale bar = 200 μm (A). Combined data from 5–6 independent experiments using 3 s separate cell strains is shown after averaging, statistical analysis and graphical representation with the bars representing the means ± SEM (control N = 6; Iso N = 5) (**P < 0.01) (B).
Mentions: To investigate wound angiogenesis in vitro, the scratch wound assay was used to determine the effect of Iso on HDMEC migration from a monolayer wound edge. β-AR activation delayed wound closure at 24 h (Fig. 3A/B). After 32 h, control wounds were 92% healed, while closure of β-AR agonist-treated wounds were significantly delayed by 25% (Fig. 3B).

Bottom Line: They play a role in wound repair but their specific role in angiogenesis is unknown.In this study, a range of in vitro assays (single cell migration, scratch wound healing, ELISAs for angiogenic growth factors and tubule formation) were performed with human dermal microvascular endothelial cells (HDMEC) to investigate and dissect mechanisms underpinning β-AR-mediated modulation of angiogenesis in chick chorioallantoic membranes (CAM) and murine excisional skin wounds. β-AR activation reduced HDMEC migration via cyclic adenosine monophosphate (cAMP)-dependent and protein kinase A (PKA)-independent mechanisms as demonstrated through use of an EPAC agonist that auto-inhibited the cAMP-mediated β-AR transduced reduction in HDMEC motility; a PKA inhibitor was, conversely, ineffective.In more complex environments, β-AR activation delayed HDMEC tubule formation and decreased angiogenesis both in the CAM assay and in murine excisional skin wounds in vivo. β-AR activation reduced HDMEC function in vitro and angiogenesis in vivo; therefore, β-AR agonists could be promising anti-angiogenic modulators in skin.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK.

Show MeSH
Related in: MedlinePlus