Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells

• Klotho is expressed in microvascular endothelial cells and dermal fibroblasts.
• Lack of Kl abrogates migration of human dermal microvascular endothelial cells.
• Lack of Kl enhances migration of human dermal fibroblasts.
• Decrease in gene expression involved in the activation of endothelial cells.

PurposeTo examine the possible role of Klotho (Kl) in human microvasculature.MethodsThe expression level of Kl in primary human dermal microvascular endothelial cells (HDMECs) and primary human dermal fibroblasts (HFb) was detected by real-time polymerase chain reaction amplification (qRT-PCR), Western blot analyses and immunohistochemistry. Migration of HDMECs and HFb was examined in monolayer wound healing “scratch assay” and Transwell assay. Proliferation of these cells was examined using Cell Proliferation BrdU incorporation assay.ResultsOur results have shown that downregulation of Kl abrogated HDMECs migration after 48 h. On the other hand, migration of HFb significantly increased after blocking Kl. Lack of Kl decreased expression of genes involved in the activation of endothelial cells and enhanced expression of genes involved in extracellular matrix remodeling and organization of connective tissue.ConclusionsThis study for the first time provides the evidence that Kl is expressed in HDMECs and HFb. Additionally, we have demonstrated that Kl is implicated in the process of angiogenesis of human dermal microvasculature.