Effects of isoprenaline on endothelial connexins and angiogenesis in a human endothelial cell culture system

Dhein, S.; Gaertner, C.; Georgieff, C.; Salameh, A.; Schlegel, F.; Mohr, F-Wilhelm.

Naunyn-Schmiedeberg's Archives of Pharmacology 388(1): 101-108

2015


ISSN/ISBN: 0028-1298
PMID: 25358823
DOI: 10.1007/s00210-014-1059-0
Accession: 052872868

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Abstract
Downregulation of endothelial connexins has been shown to result in impaired angiogenesis. Isoprenaline is known to upregulate Cx43 in cardiomyocytes. Effects of isoprenaline on endothelial connexins are unknown. We wanted to investigate whether isoprenaline might induce upregulation of connexins Cx37, Cx40, or Cx43 in human endothelial cells and whether it may promote angiogenesis. Human umbilical vein endothelial cells (HUVECs) were cultured until confluence (5 days) and subsequently seeded in Matrigel in vitro angiogenesis assays for 18 h. During the entire cell culture and angiogenesis period, cells were treated with vehicle or isoprenaline (100 nM). Finally, the resulting angiogenetic network was investigated (immuno)histologically. Moreover, expression of Cx37, Cx40, and Cx43 was determined by Western blot. In addition, we measured functional intercellular gap junction coupling by dye injection using patch clamp technique. Isoprenaline resulted in significantly enhanced expression of endothelial Cx43 and to a lower degree of Cx40 and Cx37. The number of coupling cells was significantly increased. Regarding angiogenesis, we observed significantly enhanced formation of branches and a higher complexity of the tube networks with more branches/length. Isoprenaline increases endothelial connexin expression and intercellular coupling and promotes tube formation.

Effects of isoprenaline on endothelial connexins and angiogenesis in a human endothelial cell culture system