Short Communication
Cell Biology International 10.1002/cbin.10405
The Role of eNOS in the Migration and Proliferation of Bone-Marrow Derived Endothelial Progenitor Cells and In Vitro Angiogenesis† Aizhen Lu, LiboWang, Liling Qian* Departments of Pediatrics, Children’s Hospital of Fudan University, Shanghai, 201102, P. R. China
Running title: The Role of eNOS in the Function of EPCs
Correspondence to: Liling Qian, MD, PhD Children’s Hospital of Fudan University 399 Wan Yuan Road, Shanghai 201102, P.R. China Tel: +86-21-64931913 Fax: +86-21-64931914 E-mail: [email protected]
Keywords: C-X-C Chemokine Receptor Type 4 , Endothelial progenitor cells, Endothelial nitric oxide synthase, Nitric Oxide, Nω-nitro-L-argininemethylester, Vascular Endothelial Growth Factor Receptor 2
†
This article has been accepted for publication and undergone full peer review but has not been through
the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: [10.1002/cbin.10405]
This article is protected by copyright. All rights reserved Received 19 June 2014; Revised 21 September 2014; Accepted 28 September 2014
Supported by the National Natural Science Foundation of China (No. 81270727) and Program for New Century Excellent Talents in University (NCET-12-0126)
Abstract
The role of endothelial nitric oxide synthase (eNOS) in the activities of endothelial progenitor cells (EPCs) including migration, proliferation, and tube formation in vitro was investigated. EPCs were obtained from rat bone mononuclear cells by culturing for 7-10 days in EGM-2MV and identified by their capacity for FITC-UEA-1 binding and acetylated low-density lipoprotein (Dil-ac-LDL) intake using fluorescence microscopy. Migration, proliferation and tube formation activities were assessed in the presence or absence of Nω-nitro-L-argininemethylester (L-NAME), an eNOS inhibitor. mRNA and protein expression of CXCR4, CXCR7, VEGFR2 and eNOS were detected by real-time PCR and western blotting in the presence or absence of L-NAME. Nitric oxide production was detected by nitrate reductase in the presence or absence of L-NAME. Typical spindle-shaped cells appeared on the 7th-10th day and confluence reached about 80%. The percentage of FITC-UEA-1 and Dil-ac-LDL double-stained cells was about 85%. Cell migration, proliferation, and tube formation were significantly weakened after eNOS was inhibited (P
Cell Biology International 10.1002/cbin.10405
The Role of eNOS in the Migration and Proliferation of Bone-Marrow Derived Endothelial Progenitor Cells and In Vitro Angiogenesis† Aizhen Lu, LiboWang, Liling Qian* Departments of Pediatrics, Children’s Hospital of Fudan University, Shanghai, 201102, P. R. China
Running title: The Role of eNOS in the Function of EPCs
Correspondence to: Liling Qian, MD, PhD Children’s Hospital of Fudan University 399 Wan Yuan Road, Shanghai 201102, P.R. China Tel: +86-21-64931913 Fax: +86-21-64931914 E-mail: [email protected]
Keywords: C-X-C Chemokine Receptor Type 4 , Endothelial progenitor cells, Endothelial nitric oxide synthase, Nitric Oxide, Nω-nitro-L-argininemethylester, Vascular Endothelial Growth Factor Receptor 2
†
This article has been accepted for publication and undergone full peer review but has not been through
the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: [10.1002/cbin.10405]
This article is protected by copyright. All rights reserved Received 19 June 2014; Revised 21 September 2014; Accepted 28 September 2014
Supported by the National Natural Science Foundation of China (No. 81270727) and Program for New Century Excellent Talents in University (NCET-12-0126)
Abstract
The role of endothelial nitric oxide synthase (eNOS) in the activities of endothelial progenitor cells (EPCs) including migration, proliferation, and tube formation in vitro was investigated. EPCs were obtained from rat bone mononuclear cells by culturing for 7-10 days in EGM-2MV and identified by their capacity for FITC-UEA-1 binding and acetylated low-density lipoprotein (Dil-ac-LDL) intake using fluorescence microscopy. Migration, proliferation and tube formation activities were assessed in the presence or absence of Nω-nitro-L-argininemethylester (L-NAME), an eNOS inhibitor. mRNA and protein expression of CXCR4, CXCR7, VEGFR2 and eNOS were detected by real-time PCR and western blotting in the presence or absence of L-NAME. Nitric oxide production was detected by nitrate reductase in the presence or absence of L-NAME. Typical spindle-shaped cells appeared on the 7th-10th day and confluence reached about 80%. The percentage of FITC-UEA-1 and Dil-ac-LDL double-stained cells was about 85%. Cell migration, proliferation, and tube formation were significantly weakened after eNOS was inhibited (P
