Cheng Yuan1, Lihua Ni2, Changjiang Zhang3,4,5,6, Hao Xia3, Xiaoyan Wu2. 1. Department of Gynecological Oncology, Zhongnan Hospital of Wuhan University, Wuhan, China. 2. Department of Nephrology, Zhongnan Hospital of Wuhan University, Wuhan, China. 3. Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, China. 4. Cardiovascular Research Institute, Wuhan University, Wuhan, China. 5. Hubei Key Laboratory of Cardiology, Wuhan, China. 6. Cardiovascular Disease Center, Enshi Central Hospital, Enshi, China.
Abstract
BACKGROUND: Previous study revealed that high glucose (HG) induced endothelial cell (EC) damage via endothelial-to-mesenchymal transition (EndMT). Recent studies suggested the role of Ephrin B2 in mediate ECs damage. However, the underlying mechanism remains unclear. The aim of the present study was to investigate whether Ephrin B2 mediates HG-induced EndMT in human aortic ECs (HAECs) and to determine the possible downstream signaling effector. METHODS: Primary HAECs were exposed to normal glucose (NG, 5.5 mM), HG (30 mM) and HG+Ephrin B2 small interfering RNA (siRNA), respectively. The pathological changes were investigated by light microscope and confocal microscopy. To study the effects of focal adhesion kinase (FAK) activation on Ephrin B2 in HAECs, cells were incubated with FAK siRNA in HG group. The expression of EndMT-related markers (CD31 and FSP1), Ephrin B2 and FAK were detected by qRT-PCR and western blot. RESULTS: The results showed that HG significantly inhibited the expression of CD31 and increased FSP1 compared with NG group. Moreover, Ephrin B2 was increased after HG incubation. Ephrin B2 siRNA attenuated HG-induced expression of EndMT-related markers. Furthermore, HG increased the expression of FAK and phosphorylated FAK (pho-FAK) in HAECs. In contrast, blocking Ephrin B2 could partially attenuate HG-induced FAK activation. And FAK siRNA further inhibited the EndMT-related markers in HAECs treated with HG. CONCLUSIONS: HG-induced EndMT in HAECs might be partially mediated by Ephrin B2 and the downstream FAK pathway. 2020 Cardiovascular Diagnosis and Therapy. All rights reserved.
BACKGROUND: Previous study revealed that high glucose (HG) induced endothelial cell (EC) damage via endothelial-to-mesenchymal transition (EndMT). Recent studies suggested the role of Ephrin B2 in mediate ECs damage. However, the underlying mechanism remains unclear. The aim of the present study was to investigate whether Ephrin B2 mediates HG-induced EndMT in human aortic ECs (HAECs) and to determine the possible downstream signaling effector. METHODS: Primary HAECs were exposed to normal glucose (NG, 5.5 mM), HG (30 mM) and HG+Ephrin B2 small interfering RNA (siRNA), respectively. The pathological changes were investigated by light microscope and confocal microscopy. To study the effects of focal adhesion kinase (FAK) activation on Ephrin B2 in HAECs, cells were incubated with FAK siRNA in HG group. The expression of EndMT-related markers (CD31 and FSP1), Ephrin B2 and FAK were detected by qRT-PCR and western blot. RESULTS: The results showed that HG significantly inhibited the expression of CD31 and increased FSP1 compared with NG group. Moreover, Ephrin B2 was increased after HG incubation. Ephrin B2 siRNA attenuated HG-induced expression of EndMT-related markers. Furthermore, HG increased the expression of FAK and phosphorylated FAK (pho-FAK) in HAECs. In contrast, blocking Ephrin B2 could partially attenuate HG-induced FAK activation. And FAK siRNA further inhibited the EndMT-related markers in HAECs treated with HG. CONCLUSIONS: HG-induced EndMT in HAECs might be partially mediated by Ephrin B2 and the downstream FAK pathway. 2020 Cardiovascular Diagnosis and Therapy. All rights reserved.
Entities:
Keywords:
Ephrin B2; endothelial-to-mesenchymal transition (EndMT); focal adhesion kinase (FAK); high glucose (HG); human aortic endothelial cell (HAEC)
Authors: Mélanie Héroult; Florence Schaffner; Dennis Pfaff; Claudia Prahst; Robert Kirmse; Simone Kutschera; Maria Riedel; Thomas Ludwig; Peter Vajkoczy; Ralph Graeser; Hellmut G Augustin Journal: Mol Cancer Res Date: 2010-09-13 Impact factor: 5.852