Therapeutic ultrasound protects HUVECs from ischemia/hypoxia-induced apoptosis via the PI3K-Akt pathway.

Background: Previous studies have demonstrated that therapeutic ultrasound (TUS) ameliorates angiogenesis on ischemic hind limb animals and also promotes human umbilical vein endothelial cells (HUVECs) tube formation. Apoptosis plays a key role in post-ischemic angiogenesis pathogenesis. However, the mechanisms underlying the anti-apoptotic effects of TUS are not clear. Therefore we put forward the hypothesis that TUS might promote angiogenesis during ischemia/hypoxia (I/H) by decreasing apoptosis. Methods: We investigated the cytoprotective role of TUS and the underlying mechanisms in I/H-induced HUVEC apoptosis. HUVECs were treated under hypoxic serum-starved conditions for 36 h and then treated with or without TUS (9 minutes, 1 MHz, 0.3 W/cm2). The cell viability was examined by the CCK-8 assay, apoptosis cell rate was determined by TUNEL staining and flow cytometry assay. In addition, the mitochondrial-dependent apoptosis pathway was evaluated by the protein activity of Bax, Bcl-2 and Caspase-3. Results: 1) apoptosis could be induced by I/H in HUVECs. 2) TUS attenuates HUVECs cell apoptosis induced by I/H. 3) TUS inhibits the protein expression of apoptosis modulators and effectors that regulate the mitochondrial pathway of apoptosis in HUVECs. 4) TUS increases the phosphorylation of Akt, which demonstrates the activation of the phosphoinositide 3-kinase (PI3K)- serine/threonine kinase (Akt) signal pathway. Conclusions: The present study indicates that exposure to TUS exerts a protective effect against I/H-induced apoptosis among HUVECs and that this process is mediated through the mitochondrial-dependent intrinsic apoptotic pathway. We also confirm that the PI3K-Akt signal cascade may be taken part in the TUS effects on apoptosis.