Angiogenesis effect of therapeutic ultrasound on HUVECs through activation of the PI3K-Akt-eNOS signal pathway.

Therapeutic angiogenic effects of low-intensity ultrasound have been reported in endothelial cells and animal models of hind limb ischemia. It has been shown that the proliferation, migration, and tube formation of endothelial cells play critical roles in angiogenesis. The purpose of this study was to determine the underlying mechanism of low-intensity continuous therapeutic ultrasound on angiogenesis in endothelial cells. In the present study, human umbilical vein endothelial cells (HUVECs) were simulated of low-intensity therapeutic ultrasound (TUS, 1 MHz, 0.3 W/cm(2), 9 minute per day) for 3 days, and we observed migration, tube formation, and expression of endothelial nitric oxide synthase (eNOS) and serine/threonine kinase (Akt) in HUVECs. Specific inhibitors of eNOS and phosphoinositide 3-kinase (PI3K) were added to the culture medium and TUS-induced changes in the pathways that mediate angiogenesis were investigated. After exposure to TUS, HUVECs tube formation and migration were significantly promoted, which was blocked by the eNOS inhibitor Immunofluorescence assay and Western blotting analysis demonstrated that eNOS expression in the HUVECs was significantly increased after TUS exhibition. Proteins of phosphorylated eNOS and Akt were both up-regulated after TUS stimulation. However, the specific inhibitor of PI3K not only significantly decreased the expression of p-Akt, but also down-regulated the p-eNOS. This suggested that the PI3K/Akt signal pathway might participate in modulating the activity of eNOS. In short, TUS therapy promotes angiogenesis through activation of the PI3K-Akt-eNOS signal cascade in HUVECs.

[1]  Zhi-ming Yang,et al.  Angiogenesis effect of therapeutic ultrasound on ischemic hind limb in mice. , 2014, American journal of translational research.

[2]  C. Echeverri,et al.  A chemical biology approach identified PI3K as a potential therapeutic target for neurofibromatosis type 2. , 2014, American journal of translational research.

[3]  R. Kirchmair,et al.  Low Energy Shock Wave Therapy Induces Angiogenesis in Acute Hind-Limb Ischemia via VEGF Receptor 2 Phosphorylation , 2014, PloS one.

[4]  W. Lu,et al.  Highly Sensitive and Wide-Band Tunable Terahertz Response of Plasma Waves Based on Graphene Field Effect Transistors , 2014, Scientific Reports.

[5]  Shi-ting Li,et al.  Endothelial Nitric Oxide Synthase Protects Neurons against Ischemic Injury through Regulation of Brain‐Derived Neurotrophic Factor Expression , 2014, CNS neuroscience & therapeutics.

[6]  Haibo Zhu,et al.  Protocatechuic Acid Induces Angiogenesis through PI3K‐Akt‐eNOS‐VEGF Signalling Pathway , 2013, Basic & clinical pharmacology & toxicology.

[7]  J. Mehta,et al.  ALDH2 protects against stroke by clearing 4-HNE , 2013, Cell Research.

[8]  P. Fraisl.  Crosstalk between oxygen- and nitric oxide-dependent signaling pathways in angiogenesis. , 2013, Experimental cell research.

[9]  H. Hasegawa,et al.  Low-Intensity Pulsed Ultrasound Induces Angiogenesis and Ameliorates Left Ventricular Dysfunction in a Porcine Model of Chronic Myocardial Ischemia , 2012, PloS one.

[10]  H. Gul-Uludağ,et al.  Low-intensity pulsed ultrasound-mediated stimulation of hematopoietic stem/progenitor cell viability, proliferation and differentiation in vitro , 2012, Biotechnology Letters.

[11]  Kenta Ito,et al.  Extracorporeal shock wave therapy improves the walking ability of patients with peripheral artery disease and intermittent claudication. , 2012, Circulation journal : official journal of the Japanese Circulation Society.

[12]  D. Su,et al.  Nicotinamide Postpones Stroke in Stroke‐Prone Spontaneously Hypertensive Rats , 2012, CNS neuroscience & therapeutics.

[13]  Ching‐Jen Wang,et al.  Study of vascular injuries using endothelial denudation model and the therapeutic application of shock wave: a review. , 2011, American journal of translational research.

[14]  D. O'Hare,et al.  Angiogenin induces nitric oxide synthesis in endothelial cells through PI-3 and Akt kinases. , 2010, Biochemistry.

[15]  Hao Zheng,et al.  Thymosin β4 Induces Endothelial Progenitor Cell Migration via PI3K/Akt/eNOS Signal Transduction Pathway , 2009, Journal of cardiovascular pharmacology.

[16]  R. Siegel,et al.  Ultrasound at 27 kHz increases tissue expression and activity of nitric oxide synthases in acute limb ischemia in rabbits. , 2007, Ultrasound in medicine & biology.

[17]  E. Romo,et al.  The role of nitric oxide in the post-ischemic revascularization process. , 2006, Pharmacology & therapeutics.

[18]  I. Shiojima,et al.  The HMG-CoA reductase inhibitor simvastatin activates the protein kinase Akt and promotes angiogenesis in normocholesterolemic animals. , 2000, Nature Medicine.

[19]  R. Busse,et al.  Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation , 1999, Nature.

[20]  D. Cumberland,et al.  Ultrasound enhances reporter gene expression after transfection of vascular cells in vitro. , 1999, Circulation.

[21]  Alison L. Reynolds,et al.  Targeting the PI3K/Akt/mTOR pathway in ocular neovascularization. , 2014, Advances in experimental medicine and biology.

[22]  S. Hsu,et al.  Ultrasound preexposure improves endothelial cell binding and retention on biomaterial surfaces. , 2006, Journal of biomedical materials research. Part B, Applied biomaterials.

[23]  J. Isenberg,et al.  Nitric oxide modulation of early angiogenesis , 2004, Microsurgery.