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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 decreases murine skin wound angiogenesis in vivo. Angiogenesis was assessed by analysing CD31 immunostaining of 7 μm wound sections from control and 0.1% salbutamol-treated mice 5 days post-wounding. The number of stained vessels in 10 fields of view, from one stained section per mouse, were counted in a double-blind manner and the average vessel number was calculated for both groups (N = 10–14). Arrows point to examples of CD31-positive vessels (A). Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM (** P < 0.01) (B).
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fig08: β-AR activation decreases murine skin wound angiogenesis in vivo. Angiogenesis was assessed by analysing CD31 immunostaining of 7 μm wound sections from control and 0.1% salbutamol-treated mice 5 days post-wounding. The number of stained vessels in 10 fields of view, from one stained section per mouse, were counted in a double-blind manner and the average vessel number was calculated for both groups (N = 10–14). Arrows point to examples of CD31-positive vessels (A). Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM (** P < 0.01) (B).

Mentions: To evaluate if β-AR activation altered skin wound angiogenesis, excisional murine skin wounds were treated daily with gel alone or containing 0.1% salbutamol (approximately 1.7 mM) which, at this concentration, similar to Iso, can activate all β-ARs (Baker, 2010). Salbutamol was chosen for the in vivo study as it is already widely used to treat asthma (Boskabady and Saadatinejad, 2003), therefore, should a potential clinical application be discovered, translation to the clinic could be fairly rapid through a repurposing of medicine route. Sections of murine wounds excised after 5 days were immunostained with an antibody to CD31. β-AR activation significantly reduced the number of blood vessels by an average of 56% (Fig. 8).


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 decreases murine skin wound angiogenesis in vivo. Angiogenesis was assessed by analysing CD31 immunostaining of 7 μm wound sections from control and 0.1% salbutamol-treated mice 5 days post-wounding. The number of stained vessels in 10 fields of view, from one stained section per mouse, were counted in a double-blind manner and the average vessel number was calculated for both groups (N = 10–14). Arrows point to examples of CD31-positive vessels (A). Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM (** P < 0.01) (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig08: β-AR activation decreases murine skin wound angiogenesis in vivo. Angiogenesis was assessed by analysing CD31 immunostaining of 7 μm wound sections from control and 0.1% salbutamol-treated mice 5 days post-wounding. The number of stained vessels in 10 fields of view, from one stained section per mouse, were counted in a double-blind manner and the average vessel number was calculated for both groups (N = 10–14). Arrows point to examples of CD31-positive vessels (A). Data were averaged, statistically analysed and graphically represented with the bars representing the means ± SEM (** P < 0.01) (B).
Mentions: To evaluate if β-AR activation altered skin wound angiogenesis, excisional murine skin wounds were treated daily with gel alone or containing 0.1% salbutamol (approximately 1.7 mM) which, at this concentration, similar to Iso, can activate all β-ARs (Baker, 2010). Salbutamol was chosen for the in vivo study as it is already widely used to treat asthma (Boskabady and Saadatinejad, 2003), therefore, should a potential clinical application be discovered, translation to the clinic could be fairly rapid through a repurposing of medicine route. Sections of murine wounds excised after 5 days were immunostained with an antibody to CD31. β-AR activation significantly reduced the number of blood vessels by an average of 56% (Fig. 8).

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