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Ludwig F. Zeilbeck, Verena Knobloch, Ernst R. Tamm, Andreas Ohlmann; Lithium Chloride Mediates Angiogenic Properties On Microvascular Endothelial Cells Via An Activation Of The Classical Wnt/(beta)-catenin Signaling Pathway In Vitro. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3987. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To analyze if lithium chloride (LiCl) and SB 216763, both activators of the Wnt/(beta)-catenin signaling pathway, promote angiogenic properties in cultured human dermal microvascular endothelial cells (HDMEC).
HDMEC were incubated with LiCl or SB 216763 for 3 hrs to investigate protein levels of (beta)-catenin by immunohistochemistry and western blot analyses. To examine their angiogenic potential, HDMEC were incubated with different concentrations of LiCl, SB 216763 and/or Quercetin, an inhibitor of the Wnt/(beta)-catenin pathway. Following incubation, BrdU proliferation ELISA as well as cell scratch migration assays were performed. To investigate cell survival, WST-1 assays were done after incubation of HDMEC with LiCl and SB 216763 for 3 days.
After treatment with LiCl as well as SB 216763, a substantial increase in (beta)-catenin protein level was detected by western blot analyses when compared to untreated controls. By immunohistochemistry, we observed that the increase of (beta)-catenin was followed by its translocation into the nucleus, strongly indicating that LiCl and SB 216763 can activate the Wnt/(beta)-catenin pathway in HDMEC.In comparison with untreated controls, incubation with LiCl or SB 216763 significantly increased HDMEC proliferation by 38.0 ± 0.7% (p < 0.001) or 48.0 ± 0.4% (p < 0.001) respectively. This effect was blocked when Quercetin was added. In addition, LiCl markedly increased HDMEC migration by 71.3 ± 12.3% (p < 0.05). Moreover, incubation with LiCl or SB 216763 caused an increase in HDMEC survival by 20.5 ± 2.0% (p < 0.001) or 20.0 ± 2.6% (p < 0.001).
LiCl and SB 216763 mediate angiogenic properties on cultured HDMEC, such as proliferation, migration and survival, via an activation of the classical Wnt/(beta)-catenin pathway.
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