Low-Intensity Pulsed Ultrasound Treatment Accelerates Angiogenesis by Activating YAP/TAZ in Human Umbilical Vein Endothelial Cells

Abstract

As a non-invasive method, low-intensity pulsed ultrasound (LIPUS) can accelerate fracture healing. The mechanisms responsible for the enhanced fracture healing need to be studied further. Activation of YAP/TAZ, key mediators of the Hippo signaling pathway, could promote angiogenesis and vascular remodeling. The purpose of this study was to determine whether LIPUS treatment can activate YAP/TAZ. Human umbilical vein endothelial cells (HUVEC) were used. After LIPUS treatment, Western blot and immunofluorescence staining were used for YAP/TAZ activation. Small interfering RNA (siRNA) of YAP and short hairpin LATS1/2 (shLATS1/2) were used to check whether there is cross-talk with the Hippo pathway. The phosphorylated YAP (p-127 and p-397) protein increased more than 3-fold 0.5 h after LIPUS treatment (p < 0.05). TAZ protein increased 3.0-, 2.0- and 1.5-fold 0.5, 6 and 12 h after LIPUS treatment. We found that LIPUS treatment activates YAP/TAZ, which is translocated into the cell nucleus to activate target genes. This process can be inactivated by siYAP and activated by shLATS1/2. The cross-talk with the Hippo pathway can initiate angiogenesis so as to accelerate fracture healing by LIPUS.

Keywords: Angiogenesis; LATS1/2; Low-intensity pulsed ultrasound; Nucleus translocation; YAP/TAZ.