Gender differences affect blood flow recovery in a mouse model of hindlimb ischemia

Blood flow restoration to ischemic tissue is affected by various risk factors. The aim of this study was to examine gender effects on arteriogenesis and angiogenesis in a mouse ischemic hindlimb model. C57BL/6J mice were subjected to unilateral hindlimb ischemia. Flow recovery was less and hindlimb use impairment was greater in females. No gender difference in vessel number was found at baseline, although 7 days postsurgery females had fewer α-smooth muscle actin-positive vessels in the midpoint of the adductor region. Females had higher hindlimb vascular resistance, were less responsive to vasodilators, and were more sensitive to vasoconstrictors postligation. Western blotting showed that females had higher baseline levels of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) in the calf, while 7 days postligation males had higher levels of VEGF, eNOS, and phosphorylated vasodilator stimulated phosphoprotein. Females had less angiogenesis in a Matrigel plug assay and less endothelial cell proliferation in vitro. Females have impaired recovery of flow, a finding presumably caused by multiple factors including decreased collateral remodeling, less angiogenesis, impaired vasodilator response, and increased vasoconstrictor activity; our results also suggest the possibility that new collateral formation, from capillaries, is impaired in females.

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ID: 1013 ETV-2 activated proliferation of endothelial cells and attenuated acute hindlimb ischemia in mice

Biomedical Research and Therapy ◽

10.15419/bmrat.v4is.291 ◽

2017 ◽

Vol 4 (S) ◽

pp. 87

Author(s):

Phuc Van Pham ◽

Ngoc Bich Vu ◽

Hoa Trong Nguyen ◽

Thuy Thi-Thanh Dao ◽

Ha Thi-Ngan Le ◽

...

Keyword(s):

Gene Therapy ◽

Endothelial Cells ◽

Blood Flow ◽

Viral Vector ◽

Endothelial Cell Proliferation ◽

Hindlimb Ischemia ◽

Blood Oxygen Saturation ◽

Oxygen Saturation Level ◽

Vascular Regeneration

Ischemia is the reduction of blood flow to tissues by injury of blood vessels. Depending on the sites of tissues and grade of ischemia, ischemia can cause many serious complications. This study aimed to evaluate the effects of the E-twenty six (ETS) factor Ets variant 2 (ETV2) gene expression in angiogenesis and the effect of ETV2 gene therapy in a mouse model of hindlimb ischemia. The role of ETV2 on endothelial cell proliferation was evaluated in vitro. Knockdown of ETV2 expression was done using short hairpin RNA (shRNA) lentiviral viral particles. The ETV2 viral vector was injected into the skeletal muscles at the ligated and burned sites of the hindlimb and evaluated for its efficacy as a gene therapy modality for ischemia. Vascular regeneration in mice was indirectly evaluated by changes in mouse survival, necrotic grades of the leg, normal blood oxygen saturation level (SpO2), and blood flow by trypan blue injection assay. Preliminary data showed that ETV2 expression played a role in angiogenesis of endothelial cells. ETV2 overexpression could trigger and stimulate proliferation of skeletal endothelial cells. In vivo knockdown of ETV2 expression inhibited the auto-recovery of ischemic hindlimb, while overexpression of ETV2 helped to rescue leg loss and reduce necrosis, significantly improving angiogenesis in hindlimb ischemia. Our findings demonstrate that ETV2 gene therapy is a potentially effective modality for vascular regeneration.

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Mid-luteal angiogenesis and function in the primate is dependent on vascular endothelial growth factor

Journal of Endocrinology ◽

10.1677/joe.0.1680409 ◽

2001 ◽

Vol 168 (3) ◽

pp. 409-416 ◽

Cited By ~ 50

Author(s):

SE Dickson ◽

R Bicknell ◽

HM Fraser

Keyword(s):

Vascular Endothelial Growth Factor ◽

Endothelial Cells ◽

Endothelial Cell ◽

Growth Factor ◽

Luteal Phase ◽

Smooth Muscle Actin ◽

Proliferation Index ◽

Endothelial Cell Proliferation ◽

Vascular Endothelial ◽

Endothelial Growth Factor

Vascular endothelial growth factor (VEGF) is essential for the angiogenesis required for the formation of the corpus luteum; however, its role in ongoing luteal angiogenesis and in the maintenance of the established vascular network is unknown. The aim of this study was to determine whether VEGF inhibition could intervene in ongoing luteal angiogenesis using immunoneutralisation of VEGF starting in the mid-luteal phase. In addition, the effects on endothelial cell survival and the recruitment of periendothelial support cells were examined. Treatment with a monoclonal antibody to VEGF, or mouse gamma globulin for control animals, commenced on day 7 after ovulation and continued for 3 days. Bromodeoxyuridine (BrdU), used to label proliferating cells to obtain a proliferation index, was administered one hour before collecting ovaries from control and treated animals. Ovarian sections were stained using antibodies to BrdU, the endothelial cell marker, CD31, the pericyte marker, alpha-smooth muscle actin, and 3' end DNA fragments as a marker for apoptosis. VEGF immunoneutralisation significantly suppressed endothelial cell proliferation and the area occupied by endothelial cells while increasing pericyte coverage and the incidence of endothelial cell apoptosis. Luteal function was markedly compromised by anti-VEGF treatment as judged by a 50% reduction in plasma progesterone concentration. It is concluded that ongoing angiogenesis in the mid-luteal phase is primarily driven by VEGF, and that a proportion of endothelial cells of the mid-luteal phase vasculature are dependent on VEGF support.

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Abstract 15630: Direct Renin Inhibition With Aliskiren Improves Ischemia-Induced Neovascularization

Circulation ◽

10.1161/circ.130.suppl_2.15630 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

Michel Desjarlais ◽

Sylvie Dussault ◽

Wahiba Dhahri ◽

Alain Rivard

Keyword(s):

Blood Pressure ◽

Blood Flow ◽

Renin Angiotensin System ◽

Cellular Migration ◽

High Dose ◽

Hindlimb Ischemia ◽

Adhesive Properties ◽

Tubule Formation ◽

Renin Inhibition

Background: The activation of the renin-angiotensin system is associated with impaired formation of new blood vessels (neovascularization) in response to ischemia. Aliskiren is the only direct renin inhibitor that is clinically used as an orally active antihypertensive drug. Here we tested the hypothesis that aliskiren might improve neovascularization following ischemia. Methods and Results: C57BL/6 mice were treated with a high dose of aliskiren (50 mg/kg), a low dose of aliskiren (10 mg/kg), or drinking water only. After two weeks of treatment, hindlimb ischemia was surgically induced by femoral artery removal. Treatment with aliskiren led to a significantly faster rate of blood flow recovery after hindlimb ischemia (Laser Doppler). Interestingly the lower dose of aliskiren, which did not reduce blood pressure, provided similar improvement of blood flow recuperation compared to the higher dose of aliskiren. At day 21 after surgery, Doppler flow ratios were significantly improved in mice treated with aliskiren (0.69+/-0.07 vs. 0.52+/-0.03; p<0.05). This was associated with an increased expression of angiogenic factors in ischemic muscles, including VEGF and eNOS. Endothelial progenitor cells (EPCs) have been shown to have an important role in postnatal neovascularisation. We found that aliskiren significantly increased the number of bone marrow EPCs at day 7 after ischemia (172+/-7% increase; p<0.05). Moreover, the adhesive properties of EPCs were significantly improved in mice treated with aliskiren (175+/-5% increase; p<0.05). In vitro, aliskiren improves cellular migration and tubule formation in HUVECs. This is associated with an increased expression of nitric oxide (DAF staining), and a significant reduction of oxidative stress levels (DHE staining). Importantly, the antioxidant and angiogenic properties of aliskiren in HUVECs are abolished following treatment with the NOS inhibitor L-NAME. Conclusions: Direct renin inhibition with aliskiren leads to improved ischemia-induced neovascularization that is not dependant on blood pressure lowering. The mechanisms involve beneficial effects of aliskiren on NO and angiogenic pathways in ischemic tissues, together with an increase in the number and the functional activity of EPCs.

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Hemodynamic effects of intracoronary VEGF delivery: evidence of tachyphylaxis and NO dependence of response

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1997.273.3.h1317 ◽

1997 ◽

Vol 273 (3) ◽

pp. H1317-H1323 ◽

Cited By ~ 7

Author(s):

J. J. Lopez ◽

R. J. Laham ◽

J. P. Carrozza ◽

M. Tofukuji ◽

F. W. Sellke ◽

...

Keyword(s):

Blood Flow ◽

Diastolic Pressure ◽

Rapid Development ◽

Left Ventricular ◽

No Production ◽

Vascular Endothelial ◽

Hemodynamic Effects ◽

Endothelial Growth Factor ◽

Dose Dependent Increase

Vascular endothelial growth factor (VEGF) has been utilized to improve blood flow in the setting of myocardial or peripheral vascular ischemia. In this investigation we studied the hemodynamic effects of intracoronary VEGF administration. Hemodynamic parameters and Doppler flow wire recordings from the left anterior descending coronary artery were measured after intracoronary infusion of VEGF (1, 10, and 100 micrograms) in 28 intubated pigs. Additional studies were performed using an in vitro isolated microvessel preparation. VEGF produced a highly significant dose-dependent increase in coronary blood flow (maximal 3.51 +/- 0.85-fold) in the absence of significant changes in epicardial artery diameter, a decline in mean arterial pressure (maximal 43%), and a decrease in left ventricular end-diastolic pressure (maximal 52%), all of which could be inhibited by pretreatment with NG-nitro-L-arginine. The increase in coronary flow seen with 10 or 100 micrograms VEGF was significantly greater than the maximal vasodilation achieved with serotonin or nitroglycerin and was equivalent to a maximal adenosine response. In summary, VEGF stimulates nitric oxide (NO)-dependent dilation of coronary microvessels, and repeat administrations of VEGF resulted in rapid development of tachyphylaxis to VEGF as well as serotonin, but not to nitroglycerin or adenosine, which appeared to be secondary to impaired NO production.

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The Mitogenic Effect of 17β-Estradiol on in vitro Endothelial Cell Proliferation and on in vivo Reendothelialization Are Both Dependent on Vascular Endothelial Growth Factor

Journal of Vascular Research ◽

10.1159/000025732 ◽

2000 ◽

Vol 37 (3) ◽

pp. 202-208 ◽

Cited By ~ 23

Author(s):

P. Concina ◽

S. Sordello ◽

M.A. Barbacanne ◽

R. Elhage ◽

M.T. Pieraggi ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Cell Proliferation ◽

Endothelial Cell ◽

Endothelial Cell Proliferation ◽

Vascular Endothelial ◽

Mitogenic Effect ◽

17Β Estradiol ◽

Endothelial Growth Factor

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Neural JNK3 regulates blood flow recovery after hindlimb ischemia in mice via an Egr1/Creb1 axis

Nature Communications ◽

10.1038/s41467-019-11982-4 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 4

Author(s):

Shashi Kant ◽

Siobhan M. Craige ◽

Kai Chen ◽

Michaella M. Reif ◽

Heather Learnard ◽

...

Keyword(s):

Blood Flow ◽

Growth Factors ◽

Treatment Options ◽

Clinical Manifestations ◽

Limb Ischemia ◽

The United States ◽

Hindlimb Ischemia ◽

Peripheral Ischemia ◽

Artery Disease

Abstract Diseases related to impaired blood flow such as peripheral artery disease (PAD) impact nearly 10 million people in the United States alone, yet patients with clinical manifestations of PAD (e.g., claudication and limb ischemia) have limited treatment options. In ischemic tissues, stress kinases such as c-Jun N-terminal kinases (JNKs), are activated. Here, we show that inhibition of the JNK3 (Mapk10) in the neural compartment strikingly potentiates blood flow recovery from mouse hindlimb ischemia. JNK3 deficiency leads to upregulation of growth factors such as Vegfa, Pdgfb, Pgf, Hbegf and Tgfb3 in ischemic muscle by activation of the transcription factors Egr1/Creb1. JNK3 acts through Forkhead box O3 (Foxo3a) to suppress the activity of Egr1/Creb1 transcription regulators in vitro. In JNK3-deficient cells, Foxo3a is suppressed which leads to Egr1/Creb1 activation and upregulation of downstream growth factors. Collectively, these data suggest that the JNK3-Foxo3a-Egr1/Creb1 axis coordinates the vascular remodeling response in peripheral ischemia.

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VEGF-A-Cleavage by FSAP and Inhibition of Neo-Vascularization

Cells ◽

10.3390/cells8111396 ◽

2019 ◽

Vol 8 (11) ◽

pp. 1396 ◽

Cited By ~ 1

Author(s):

Özgür Uslu ◽

Joerg Herold ◽

Sandip M. Kanse

Keyword(s):

Growth Factor ◽

Tissue Injury ◽

Factor Vii ◽

Endothelial Cell Proliferation ◽

Vegf Receptor ◽

Vascular Endothelial ◽

The Matrix ◽

Endothelial Growth Factor

Alternative splicing leads to the secretion of multiple forms of vascular endothelial growth factor-A (VEGF-A) that differ in their activity profiles with respect to neovascularization. FSAP (factor VII activating protease) is the zymogen form of a plasma protease that is activated (FSAPa) upon tissue injury via the release of histones. The purpose of the study was to determine if FSAPa regulates VEGF-A activity in vitro and in vivo. FSAP bound to VEGF165, but not VEGF121, and VEGF165 was cleaved in its neuropilin/proteoglycan binding domain. VEGF165 cleavage did not alter its binding to VEGF receptors but diminished its binding to neuropilin. The stimulatory effects of VEGF165 on endothelial cell proliferation, migration, and signal transduction were not altered by FSAP. Similarly, proliferation of VEGF receptor-expressing BAF3 cells, in response to VEGF165, was not modulated by FSAP. In the mouse matrigel model of angiogenesis, FSAP decreased the ability of VEGF165, basic fibroblast growth factor (bFGF), and their combination, to induce neovascularization. Lack of endogenous FSAP in mice did not influence neovascularization. Thus, FSAP inhibited VEGF165-mediated angiogenesis in the matrigel model in vivo, where VEGF’s interaction with the matrix and its diffusion are important.

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Cathepsin K Deficiency Impaired Ischemia-Induced Neovascularization in Aged Mice

Stem Cells International ◽

10.1155/2020/6938620 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Xueling Yue ◽

Haiying Jiang ◽

Ying Xu ◽

Manli Xia ◽

Xian-Wu Cheng

Keyword(s):

Blood Flow ◽

Flow Analysis ◽

Cathepsin K ◽

Cellular Functions ◽

Aged Mice ◽

Blood Flow Ratio ◽

K Deficiency ◽

Signaling Activation ◽

Endothelial Growth Factor

Background. Aging is a major risk factor for cardiovascular disease. Cysteine protease cathepsin K (CatK) has been implicated in the process of angiogenesis, but the exact roles of individual CatK in vessel formation during aging are poorly understood. Methods and Results. To study the putative role of CatK in ischemia-induced angiogenesis, we applied a hindlimb ischemia model to aged wild-type (CatK+/+) and CatK-deficient (CatK−/−) mice. A serial laser Doppler blood-flow analysis revealed that the recovery of the ischemic/normal blood-flow ratio in the aged CatK−/−mice was impaired throughout the follow-up period. On postoperative day 14, CatK deficiency had also impaired capillary formation. CatK deficiency reduced the levels of cleaved Notch1, phospho-Akt, and/or vascular endothelial growth factor (VEGF) proteins in the ischemic muscles and bone marrow-derived c-Kit+ cells. A flow cytometry analysis revealed that CatK deficiency reduced the numbers of endothelial progenitor cell (EPC)-like CD31+/c-Kit+ cells in the peripheral blood as well as the ischemic vasculature. In vitro experiments, CatK−/− impaired bone-derived c-Kit+ cellular functions (migration, invasion, proliferation, and tubulogenesis) in aged mice. Our findings demonstrated that aging impaired the ischemia-induced angiogenesis associated with the reductions of the production and mobilization of CD31+/c-Kit+ cells in mice. Conclusions. These findings established that the impairment of ischemia-induced neovascularization in aged CatK−/− mice is due, at least in part, to the reduction of EPC mobilization and the homing of the cells into vasculature that is associated with the impairment of Notch1 signaling activation at advanced ages.

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