Abstract 1196: Vascular Endothelial Cells specific Endothelin-1 mediates Early Inflammation in Murine Model of Neointimal Formation

Complex mechanism of the chronic inflammatory process in vascular injury and atherosclerosis involves various types of cells and cytokines. Previous in vitro studies suggest that Endothelin-1 (ET-1) is produced by those cells, induces expression of several adhesion molecules and mediates the leukocyte-endothelial interaction. In the present study, we sought to determine whether local endothelial ET-1 plays a direct effect on early inflammation and neointimal formation as a response to vascular injury in vivo . Using Cre-loxP system and Tie-2 Cre promoter, Vascular Endothelial ET-1 Knockout (VEETKO) mice was generated. The mice showed 60% reduction of ET-1 mRNA level at major organs and undetected serum ET-1 level. To induce vascular inflammation, complete ligation of the left common carotid artery was performed in 12-weeks-old male VEETKO (n=35) and Wild Type (WT) littermates (n=34). Inhibition of vascular endothelial ET-1 prevented the 4-fold increase in ET-1 mRNA level of carotid artery after ligation, in contrast to that of WT mice. This prevention resulted in lower vascular inflammation revealed by lower mRNA and protein level of PECAM-1, ICAM-1, and MCP-1 in VEETKO mice. The lack of ET-1 in endothelium further inhibited the adhesion of inflammatory cells at early days after ligation as compared to that adhered in endothelium of WT mice (cell numbers, 20.7±4.9 vs. 54.6±3.9 respectively, p<0.05, VEEETKO n=8/WT n=10). Observation at 4 weeks after ligation noted lower neointimal formation in VEETKO mice (n=11) as compared to WT littermates (n=12) (neointimal/medial ratio, 0.2±0.03 vs. 0.8±0.25 respectively, p<0.05). Changes in carotid flow gave no hemodynamical effect as sistolic blood pressure maintained significantly lower in VEETKO mice as compared to WT mice (105.7±1.4 vs. 115.9±1.6 mmHg before ligation, p<0.05 and 105.7±2.4 vs.119.08±1.08 mmHg after ligation, p<0.05 n=20 each). In conclusion, these data demonstrated that lack of vascular endothelial ET-1 reduced inflammatory response of the vascular wall followed by inhibition of neointimal formation confirming the role of local endothelial ET-1 in mediating inflammation in vascular injury, in addition to its potent vasoconstrictor and proliferating effect.

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miR-24 limits aortic vascular inflammation and murine abdominal aneurysm development

Nature Communications ◽

10.1038/ncomms6214 ◽

2014 ◽

Vol 5 (1) ◽

Cited By ~ 104

Author(s):

Lars Maegdefessel ◽

Joshua M. Spin ◽

Uwe Raaz ◽

Suzanne M. Eken ◽

Ryuji Toh ◽

...

Keyword(s):

Vascular Endothelial Cells ◽

Vascular Inflammation ◽

Aortic Tissue ◽

Vascular Endothelial ◽

Vascular Medicine ◽

Abdominal Aneurysm ◽

Abdominal Aortic ◽

Smooth Muscle Cell Migration

Abstract Identification and treatment of abdominal aortic aneurysm (AAA) remain among the most prominent challenges in vascular medicine. MicroRNAs (miRNAs) are crucial regulators of cardiovascular pathology and represent intriguing targets to limit AAA expansion. Here we show, by using two established murine models of AAA disease along with human aortic tissue and plasma analysis, that miR-24 is a key regulator of vascular inflammation and AAA pathology. In vivo and in vitro studies reveal chitinase 3-like 1 (Chi3l1) to be a major target and effector under the control of miR-24, regulating cytokine synthesis in macrophages as well as their survival, promoting aortic smooth muscle cell migration and cytokine production, and stimulating adhesion molecule expression in vascular endothelial cells. We further show that modulation of miR-24 alters AAA progression in animal models, and that miR-24 and CHI3L1 represent novel plasma biomarkers of AAA disease progression in humans.

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Synthesis and physiological activity of heterodimers comprising different splice forms of vascular endothelial growth factor and placenta growth factor

Biochemical Journal ◽

10.1042/bj3160703 ◽

1996 ◽

Vol 316 (3) ◽

pp. 703-707 ◽

Cited By ~ 34

Author(s):

Ralf BIRKENHÄGER ◽

Bernard SCHNEPPE ◽

Wolfgang RÖCKL ◽

Jörg WILTING ◽

Herbert A. WEICH ◽

...

Keyword(s):

Endothelial Cells ◽

Growth Factor ◽

Vascular Endothelial Cells ◽

Physiological Activity ◽

Expression System ◽

Vascular Endothelial ◽

Placenta Growth Factor ◽

Splice Forms

Vascular endothilial growth factor (VEGF) and placenta growth factor (PIGF) are members of a dimeric-growth-factor family with angiogenic properties. VEGF is a highly potent and specific mitogen for endothelial cells, playing a vital role in angiogenesis in vivo. The role of PIGF is less clear. We expressed the monomeric splice forms VEGF-165, VEGF-121, PIGF-1 and PlGF-2 as unfused genes in Escherichia coli using the pCYTEXP expression system. In vitro dimerization experiments revealed that both homo- and hetero-dimers can be formed from these monomeric proteins. The dimers were tested for their ability to promote capillary growth in vivo and stimulate DNA synthesis in cultured human vascular endothelial cells. Heterodimers comprising different VEGF splice forms, or combinations of VEGF/PlGF splice forms, showed mitogenic activity. The results demonstrate that four different heterodimeric growth factors are likely to have as yet uncharacterized functions in vivo.

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Angiopoietin-related growth factor (AGF) promotes angiogenesis

Blood ◽

10.1182/blood-2003-04-1272 ◽

2004 ◽

Vol 103 (10) ◽

pp. 3760-3765 ◽

Cited By ~ 62

Author(s):

Yuichi Oike ◽

Yasuhiro Ito ◽

Hiromitsu Maekawa ◽

Tohru Morisada ◽

Yoshiaki Kubota ◽

...

Keyword(s):

Growth Factor ◽

Transgenic Mice ◽

Vascular Endothelial Cells ◽

Direct Injection ◽

Angiogenic Factor ◽

Epidermal Keratinocytes ◽

Vascular Endothelial ◽

Novel Therapeutic Strategies

Abstract We report here the identification of angiopoietin-related growth factor (AGF) as a positive mediator for angiogenesis. To investigate the biologic function of AGF in angiogenesis, we analyzed the vasculature in the dermis of transgenic mice expressing AGF in mouse epidermal keratinocytes (K14-AGF). K14-AGF transgenic mice were grossly red, especially in the ears and snout, suggesting that hypervascularization had occurred in their skin. Histologic examination of ear skin from K14-AGF transgenic mice revealed increased numbers of microvessels in the dermis, whereas the expression of several angiogenic factors, such as basic fibroblast growth factor (bFGF), vascular endothelial growth factors (VEGFs), and angiopoietin-1 (Ang-1), was decreased. We showed that AGF is a secreted protein and does not bind to tyrosine kinase with immunoglobulin and EGF-homology domain (Tie1) or Tie2 receptors. An in vitro chamber assay revealed that AGF directly promotes chemotactic activity of vascular endothelial cells. Both mouse corneal and matrigel plug assays showed that AGF induces neovascularization in vivo. Furthermore, we found that plasma leakage occurred after direct injection of AGF into the mouse dermis, suggesting that AGF directly induces a permeability change in the local vasculature. On the basis of these observations, we propose that AGF is a novel angiogenic factor and that handling of its biologic functions could lead to novel therapeutic strategies for control of angiogenesis.

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Interaction between vascular endothelial cells and vascular intimal spindle-shaped cells in vitro

Journal of Cell Science ◽

10.1242/jcs.60.1.89 ◽

1983 ◽

Vol 60 (1) ◽

pp. 89-102

Author(s):

D de Bono ◽

C. Green

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Vascular Endothelial Cells ◽

Three Dimensional ◽

Vascular Endothelial ◽

Pure Cultures ◽

Species Specific ◽

Endothelial Growth Factor

The interactions between human or bovine vascular endothelial cells and fibroblast-like vascular intimal spindle-shaped cells have been studied in vitro, using species-specific antibodies to identify the different components in mixed cultures. Pure cultures of endothelial cells grow as uniform, nonoverlapping monolayers, but this growth pattern is lost after the addition of spindle cells, probably because the extracellular matrix secreted by the latter causes the endothelial cells to modify the way they are attached to the substrate. The result is a network of tubular aggregates of endothelial cells in a three-dimensional ‘polylayer’ of spindle-shaped cells. On the other hand, endothelial cells added to growth-inhibited cultures of spindle-shaped cells will grow in sheets over the surface of the culture. Human endothelial cells grown in contact with spindle-shaped cells have a reduced requirement for a brain-derived endothelial growth factor. The interactions of endothelial cells and other connective tissue cells in vitro may be relevant to the mechanisms of endothelial growth and blood vessel formation in vivo, and emphasize the potential importance of extracellular matrix in controlling endothelial cell behaviour.

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EDRF as a possible mediator of sepsis-induced arteriolar dilation in skeletal muscle

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1992.262.3.h880 ◽

1992 ◽

Vol 262 (3) ◽

pp. H880-H887 ◽

Cited By ~ 2

Author(s):

A. S. Lubbe ◽

R. N. Garrison ◽

H. M. Cryer ◽

N. L. Alsip ◽

P. D. Harris

Keyword(s):

Skeletal Muscle ◽

Cardiac Output ◽

Defense Mechanisms ◽

Vascular Endothelial Cells ◽

Early Period ◽

Vascular Endothelial ◽

Low Cardiac Output ◽

E Coli

Vascular endothelial cells influence microvessel diameters in vivo and in vitro and participate in host-defense mechanisms during sepsis. We examined whether small arteriole dilation in skeletal muscle during high cardiac output bacteremia (HOB) and low cardiac output live Escherichia coli sepsis (LOS) is mediated by an endothelium-derived relaxing factor (EDRF). Local chemical blockade of EDRF by hydroquinone (HQ) substantially blunted acetylcholine-induced dilation of small arterioles. HQ also prevented large arteriole (55-135 microns) constriction and small arteriole (6-22 microns) dilation in the cremaster muscle of rats during HOB. In LOS, small arteriole dilation was also prevented by HQ but only during the early period when blood pressure was unchanged from baseline. HQ did not alter large arteriole constriction during LOS. We conclude that small arteriole vasodilation in skeletal muscle is mediated at least in part by EDRF during bacteremia. Because EDRF cannot mediate large arteriole constriction and because HQ blunted large arteriole constriction during HOB, we now suspect that HQ also interferes at least in part with some large arteriole vasoconstrictor mechanism, possibly leukotrienes or an endothelium-derived constricting factor, which mediates large arteriole constriction during HOB. Our data also suggest that large arteriole constriction during LOS is partly mediated by factors that are unaffected by HQ. The endothelium appears to play an important role in the microcirculatory responses of skeletal muscle to live E. coli sepsis through more than one mechanism.

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A NEW BUTADIENE DERIVATIVE, T-686, INHIBITS PLASMINOGEN ACTIVATOR INHIBITOR TYPE-1 PRODUCTION IN VITRO BY CULTURED HUMAN VASCULAR ENDOTHELIAL CELLS AND DEVELOPMENT OF ATHEROSCLEROTIC LESIONS IN VIVO IN RABBITS

Thrombosis Research ◽

10.1016/s0049-3848(97)00016-9 ◽

1997 ◽

Vol 85 (4) ◽

pp. 305-314 ◽

Cited By ~ 6

Author(s):

Boris Vinogradsky ◽

Stephen P Bell ◽

Janet Woodcock-Mitchell ◽

Akio Ohtani ◽

Satoshi Fujii

Keyword(s):

Vascular Endothelial Cells ◽

Plasminogen Activator Inhibitor ◽

Vascular Endothelial ◽

Atherosclerotic Lesions ◽

Inhibitor Type ◽

Activator Inhibitor Type ◽

Activator Inhibitor

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CYLD regulates angiogenesis by mediating vascular endothelial cell migration

Blood ◽

10.1182/blood-2009-10-248526 ◽

2010 ◽

Vol 115 (20) ◽

pp. 4130-4137 ◽

Cited By ~ 52

Author(s):

Jinmin Gao ◽

Lei Sun ◽

Lihong Huo ◽

Min Liu ◽

Dengwen Li ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Migration ◽

Endothelial Cell ◽

Vascular Endothelial Cells ◽

Vascular Endothelial Cell ◽

Tube Formation ◽

Endothelial Cell Migration ◽

Vascular Endothelial

Cylindromatosis (CYLD) is a deubiquitinase that was initially identified as a tumor suppressor and has recently been implicated in diverse normal physiologic processes. In this study, we have investigated the involvement of CYLD in angiogenesis, the formation of new blood vessels from preexisting ones. We find that knockdown of CYLD expression significantly impairs angiogenesis in vitro in both matrigel-based tube formation assay and collagen-based 3-dimensional capillary sprouting assay. Disruption of CYLD also remarkably inhibits angiogenic response in vivo, as evidenced by diminished blood vessel growth into the angioreactors implanted in mice. Mechanistic studies show that CYLD regulates angiogenesis by mediating the spreading and migration of vascular endothelial cells. Silencing of CYLD dramatically decreases microtubule dynamics in endothelial cells and inhibits endothelial cell migration by blocking the polarization process. Furthermore, we identify Rac1 activation as an important factor contributing to the action of CYLD in regulating endothelial cell migration and angiogenesis. Our findings thus uncover a previously unrecognized role for CYLD in the angiogenic process and provide a novel mechanism for Rac1 activation during endothelial cell migration and angiogenesis.

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Origin and diversity of embryonic endothelium/endocardium

10.1093/med/9780198757269.003.0005 ◽

2018 ◽

Author(s):

LeShana SaintJean ◽

H.S. Baldwin

Keyword(s):

Endothelial Cells ◽

Heart Development ◽

Vascular Endothelial Cells ◽

Coronary Vessels ◽

Cell Types ◽

Genetic Profile ◽

Vascular Endothelial ◽

Tremendous Progress

The endocardium represents a distinct population of endothelial cells that arises during the initiation of heart development. Endocardial cells can easily be distinguished from most of the other cardiac cell types. However, endocardial and vascular endothelial cells contain a similar genetic profile that limits the ability to study each group independently. Despite these limitations, tremendous progress has been made in identifying the different roles of endocardial cells throughout heart development. Initial studies focused on the origin of endocardial cells and their role in valvulogenesis, trabeculation, and formation of the ventricular and atrial septum. With the advancement of microscopy and the availability of endocardial specific reporter models (in vitro and in vivo) we have obtained more insight into the molecular, structural, and functional complexity of the endocardium. Additional studies have demonstrated how the endocardium is also involved in the development of coronary vessels within the compact myocardium and in heart regeneration.

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Adipose tissue-derived stem cells inhibit neointimal formation in a paracrine fashion in rat femoral artery

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00391.2009 ◽

2010 ◽

Vol 298 (2) ◽

pp. H415-H423 ◽

Cited By ~ 31

Author(s):

Masao Takahashi ◽

Etsu Suzuki ◽

Shigeyoshi Oba ◽

Hiroaki Nishimatsu ◽

Kenjiro Kimura ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Femoral Artery ◽

Vascular Endothelial Cells ◽

Vascular Endothelial ◽

Neointimal Formation ◽

Paracrine Factors ◽

Endothelial Layer ◽

Endothelial Repair

Subcutaneous adipose tissue contains a lot of stem cells [adipose-derived stem cells (ASCs)] that can differentiate into a variety of cell lineages. In this study, we isolated ASCs from Wistar rats and examined whether ASCs would efficiently differentiate into vascular endothelial cells (ECs) in vitro. We also administered ASCs in a wire injury model of rat femoral artery and examined their effects. ASCs expressed CD29 and CD90, but not CD34, suggesting that ASCs resemble bone marrow-derived mesenchymal stem cells. When induced to differentiate into ECs with endothelial growth medium (EGM), ASCs expressed Flt-1, but not Flk-1 or mature EC markers such as CD31 and vascular endothelial cadherin. ASCs produced angiopoietin-1 when they were cultured in EGM. ASCs stimulated the migration of EC, as assessed by chemotaxis assay. When ASCs that were cultured in EGM were injected in the femoral artery, the ASCs potently and significantly inhibited neointimal formation without being integrated in the endothelial layer. EGM-treated ASCs significantly suppressed neointimal formation even when they were administered from the adventitial side. ASC administration significantly promoted endothelial repair. These results suggested that although ASCs appear to have little capacity to differentiate into mature ECs, ASCs have the potential to secrete paracrine factors that stimulate endothelial repair. Our results also suggested that ASCs inhibited neointimal formation via their paracrine effect of stimulation of EC migration in situ rather than the direct integration into the endothelial layer.

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