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Updated: Jan 18, 2026

HUVEC Tube-formation Assay to Evaluate the Impact of Natural Products on Angiogenesis
Published on: June 24, 2019
Twelve-week nitrate-rich beetroot extract supplementation improves lower limb vascular function and serum angiogenic
Thiago Silveira Alvares1, Vivian Pinheiro1, David N Proctor2
1Multidisciplinary Center UFRJ-Macaé, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
Abstract:
Postmenopausal women have reduced nitric oxide (NO) bioavailability and impairments within the microcirculation and lower limb conduit arteries, key factors contributing to cardiovascular disease (CVD) development and mortality. Dietary nitrate supplementation has shown promise in improving vascular function, yet most studies are limited to short-term interventions and upper-limb conduit artery function assessment. We investigated whether a 12-wk nitrate-rich beetroot extract supplementation improves lower-limb conduit artery and skeletal muscle microvascular function, alongside serum angiogenic potential, in postmenopausal women. Postmenopausal women (60-85 yr) were randomized to consume nitrate-rich (8.8 mmol nitrate, n = 8) or nitrate-depleted beetroot extract (0.7 mmol nitrate, n = 8) daily for 12 wk. Femoral artery endothelial function was assessed via flow-mediated dilation (FMD), whereas tibialis anterior muscle microvascular reactivity was evaluated by near-infrared spectroscopy-derived reperfusion slope (StO2 slope 2) every 4 wk. Serum angiogenic potential was evaluated through endothelial tube formation assays using human umbilical vein endothelial cells. Serum nitrate increased significantly at weeks 4, 8, and 12 in the nitrate-rich group (P < 0.05), with no changes in controls. Femoral artery FMD improved significantly after 12 wk (P < 0.05), whereas tibialis anterior microvascular reactivity increased after 8 and 12 wk (P < 0.05). Serum from nitrate-supplemented participants at week 12 induced a 1.8-fold increase in vitro tube formation compared with baseline (P < 0.05). Collectively, the current findings suggest nitrate-rich beetroot extract as a potential dietary strategy to attenuate the vascular detrimental effects of menopause and reduce CVD risk in older women.NEW & NOTEWORTHY Postmenopausal women exhibit impaired lower limb vascular function and reduced angiogenic capacity, increasing cardiovascular risk. This study demonstrates that 12 wk of nitrate-rich beetroot extract supplementation enhances femoral artery endothelial function, skeletal muscle microvascular reactivity, and serum angiogenic potential, offering a promising dietary strategy to improve vascular health in this population.
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