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.
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Jing-Juan
JJ
Huang
Department of Vascular Surgery, Ren Ji Hospital, Shanghai Jiaotong University School of Medicine Dongfang Road 1630, Shanghai 200127, China.
Yi-Qin
YQ
Shi
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Rui-Lin
RL
Li
Department of Otolaryngology, Gong li Hospital Miaopu Road 219, Shanghai 200135, China.
An
A
Hu
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Zhao-Yang
ZY
Lu
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Liang
L
Weng
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Shen-Qi
SQ
Wang
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Yi-Peng
YP
Han
Department of Vascular Surgery, Ren Ji Hospital, Shanghai Jiaotong University School of Medicine Dongfang Road 1630, Shanghai 200127, China.
Lan
L
Zhang
Department of Cardiology, Shanxi Cardiovascular Hospital Yifen Street 18, Taiyuan 030024, China.
Bao
B
Li
Department of Vascular Surgery, Ren Ji Hospital, Shanghai Jiaotong University School of Medicine Dongfang Road 1630, Shanghai 200127, China.
Chang-Ning
CN
Hao
Department of Gerontology, XinHua Hospital, Shanghai Jiaotong University School of Medicine Kongjiang Road 1665, Shanghai 200092, China.
Jun-Li
JL
Duan
Therapeutic ultrasound; angiogenesis; human umbilical vein endothelial cells; migration