An Assay to Measure Angiogenesis in Human Fat Tissue

Greenway, Frank L.; Liu, Zhijun; Yu, Ying; Caruso, Mary K.; Roberts, Andrew T.; Lyons, John; Schwimer, Joshua E.; Gupta, Alok; Bellanger, Drake E.; Guillot, Thomas S.; Woltering, Eugene A.

doi:10.1007/s11695-007-9089-z

Cited by 15 publications

(13 citation statements)

References 12 publications

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“…Early studies described a stimulation of angiogenesis by adipose tissue in animal models, such as grafts of rabbit adipose tissue onto cornea (13) or onto the brain surface (14). A more recent study showed de novo formation of vessels from human adipose tissue embedded in fibrin thrombin clot (25). The adipose tissue samples that we obtained from obese subjects induced a powerful angiogenic response when layered on CAM, a well-known and robust model of angiogenesis (27).…”

Section: Discussionmentioning

confidence: 78%

“…Hypoxia and inflammation strongly promote angiogenesis (2,3,25,32). It was shown in mice that oxygen supply is decreased in hypertrophic adipose tissue (41), and expression and macrophage density of cytokines in adipose tissue are increased in obesity (37,42).…”

Section: Discussionmentioning

confidence: 99%

“…The characteristics of adipose tissue that could influence angiogenesis in humans, such as whether the tissue is of subcutaneous or visceral origin, the presence of infiltrating macrophages, and the profile of expression of angiogenic factors are still unknown. Very few studies (8,24,25) have tried to inhibit angiogenic factors to identify their role in adipose tissue angiogenesis.…”

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confidence: 99%

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Angiogenesis Associated With Visceral and Subcutaneous Adipose Tissue in Severe Human Obesity

et al. 2008

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OBJECTIVE— The expansion of adipose tissue is linked to the development of its vasculature. However, the regulation of adipose tissue angiogenesis in humans has not been extensively studied. Our aim was to compare the angiogenesis associated with subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) from the same obese patients in an in vivo model. RESEARCH DESIGN AND METHODS— Adipose tissue samples from visceral (VAT) and subcutaneous (SAT) sites, obtained from 36 obese patients (mean BMI 46.5 kg/m 2 ) during bariatric surgery, were layered on chick chorioallantoïc membrane (CAM). RESULTS— Both SAT and VAT expressed angiogenic factors without significant difference for vascular endothelial growth factor (VEGF) expression. Adipose tissue layered on CAM stimulated angiogenesis. Angiogenic stimulation was macroscopically detectable, with engulfment of the samples, in 39% and was evidenced by angiography in 59% of the samples. A connection between CAM and adipose tissue vessels was evidenced by immunohistochemistry, with recruitment of both avian and human endothelial cells. The angiogenic potency of adipose tissue was not related to its localization (with an angiogenic stimulation in 60% of SAT samples and 61% of VAT samples) or to adipocyte size or inflammatory infiltrate assessed in adipose samples before the graft on CAM. Stimulation of angiogenesis by adipose tissue was nearly abolished by bevacizumab, which specifically targets human VEGF. CONCLUSIONS— We have established a model to study the regulation of angiogenesis by human adipose tissue. This model highlighted the role of VEGF in angiogenesis in both SAT and VAT.

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Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Angiogenesis Associated With Visceral and Subcutaneous Adipose Tissue in Severe Human Obesity

et al. 2008

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“…18 Subcutaneous adipose tissue was removed from the anterior abdominal area of morbidly obese subjects undergoing gastric bypass surgery through the incisions placed to insert the laparoscopic instruments. The fat was placed directly into sterile assay media (see below), transported directly to the laboratory from the surgery in the sterile container and processed under a laminar flow hood.…”

Section: Methodsmentioning

confidence: 99%

“…The angiogenesis media were replaced every 48 h and appropriate concentrations of fresh enterostatin were added. Wells were evaluated for the angiogenic response as described by Greenway et al 18 An observer unbiased to the treatment protocols evaluated the angiogenic response using a semi-quantitative visual rating scale. 19 Essentially, the tissue was viewed under an inverted microscope, visually divided into four quadrants and each quadrant was given a numeric score from 0 to 4 based on the neovessel's length, density and percentage of the quadrants' circumference involved with the angiogenic response.…”

Section: Methodsmentioning

confidence: 99%

Enterostatin inhibition of angiogenesis: possible role of pAMPK and vascular endothelial growth factor A (VEGF-A)

Park

Lyons

et al. 2008

Int J Obes

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Introduction:We utilized a genomic analysis of the response of neuronal GT1-7 cells to enterostatin to identify pathways responsive to this peptide. This information, together with reported properties of the enterostatin receptor, suggested that enterostatin may have an effect on angiogenesis. Method: To investigate this hypothesis, we studied the effect of enterostatin as an antiangiogenic agent in two angiogenic tissue culture model systems.Results: Enterostatin induced a 50% or greater inhibition in the angiogenic response of human fat cells and had a U-shaped bimodal dose-response effect in inhibiting angiogenesis in a human placental vein angiogenesis model. To further understand this response, we tested enterostatin's effect in a human hepatoma cell line (HepG2 cells) that was subjected to glucose deprivation, a condition known to induce angiogenesis in other tumor cell lines. Phosphorylated AMP kinase (pAMPK) levels and vascular endothelial growth factor A (VEGF-A) mRNA expression were elevated robustly after incubation of HepG2 cells in the absence of glucose for 4 h, but 15 min incubation with enterostatin dramatically inhibited this pAMPK activation and reduced VEGF-A gene expression after 1 h incubation with enterostatin. The AMPK activator 5-aminoimidazole-4-carboximide ribonucleoside (AICAR) induced VEGF-A expression. Summary: These data suggest that enterostatin has an antiangiogenic effect and suggest that it regulates VEGF-A gene expression through inhibition of AMPK activity.

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α‐Tocopheryl phosphate—An activated form of vitamin E important for angiogenesis and vasculogenesis?

2012

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Vitamin E was originally discovered as a dietary factor essential for reproduction in rats. Since then, vitamin E has revealed many important molecular properties such as the scavenging of reactive oxygen and nitrogen species or the modulation of signal transduction and gene expression in antioxidant and nonantioxidant manners. A congenital disease, ataxia with vitamin E deficiency, which is characterized by impaired enrichment of α-tocopherol (αT) in plasma due to mutations in the α-tocopherol transfer protein gene, has been discovered. An effect of vitamin E on angiogenesis and vasculogenesis has been observed in several studies, and recently, it has been demonstrated in the placenta of pregnant ewes, possibly involving the stimulation of vascular endothelial growth factor (VEGF) expression. We recently observed that the phosphorylated form of αT, α-tocopheryl phosphate (αTP), increases the expression of VEGF. We propose that the stimulatory effect of αT on angiogenesis and vasculogenesis is potentiated by phosphorylation to αTP, which may act as a cofactor or active lipid mediator increasing VEGF expression. Increased VEGF expression and consequent enhanced angiogenesis and vasculogenesis induced by αTP may explain not only the essential roles of vitamin E on reproduction, but also its beneficial effects against pre-eclampsia, ischemia/reperfusion injury, and during wound healing. It may also serve as a survival factor for brain and muscle cells. The finding that αTP may regulate vasculogenesis may indicate potential, important pathophysiological implications.

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An Assay to Measure Angiogenesis in Human Fat Tissue

Cited by 15 publications

References 12 publications

Angiogenesis Associated With Visceral and Subcutaneous Adipose Tissue in Severe Human Obesity

Angiogenesis Associated With Visceral and Subcutaneous Adipose Tissue in Severe Human Obesity

Enterostatin inhibition of angiogenesis: possible role of pAMPK and vascular endothelial growth factor A (VEGF-A)

α‐Tocopheryl phosphate—An activated form of vitamin E important for angiogenesis and vasculogenesis?

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