Low‐dose aspirin promotes endothelial progenitor cell migration and adhesion and prevents senescence

Abstract

Circulating endothelial progenitor cells (EPCs) play a key role in restoring endothelial function and enhancing angiogenesis. However, the effects of low-dose aspirin on circulating EPCs are not well known. We investigated the effects of low-dose aspirin on EPC migration, adhesion, senescence, proliferation, apoptosis and endothelial nitric oxide synthase (eNOS) expression. EPC migration was detected by a modified Boyden chamber assay. EPC adhesion assay was performed by counting adherent cells on fibronectin-coated culture dishes. EPC senescence was assessed by both senescence-associated-beta-galactosidase staining and DAPI staining. EPC proliferation was analyzed by MTT assay. EPC apoptosis was evaluated by flow cytometric analysis. eNOS protein expression was measured by Western blotting analysis. Aspirin promoted EPC migratory and adhesive capacity at concentrations between 0.1 and 100micromol/L and prevented senescence at concentrations between 50 and 100micromol/L. Meanwhile, aspirin in a range of these concentrations did not affect EPC proliferation, apoptosis or eNOS expression. Our findings indicate that low-dose aspirin promotes migration and adhesion and delays the onset of senescence of EPCs.

Sulfhydryl Angiotensin-Converting Enzyme Inhibitor Promotes Endothelial Cell Survival through Nitric-Oxide Synthase, Fibroblast Growth Factor-2, and Telomerase Cross-Talk

“…The mechanism whereby ACEI promote TERT, in the present conditions, seems to be related to the activation of the FGF-2 pathway, because its silencing or neutralization abrogates TERT expression. Although the evidence indicating down-regulation of telomerase by oxidative stress is overwhelming (Haendeler et al, 2004;Voghel et al, 2008;Erusalimsky and Skene, 2009), interventions that favor its up-regulation are limited to NO donor drugs (Vasa et al, 2000), statins (Haendeler et al, 2004), low-dose aspirin (Hu et al, 2008), estrogen (Grasselli et al, 2008), and FGF-2 (Kurz et al, 2003). The direct stimulation of human TERT expression by NO, either endogenously produced or delivered by NO donor agents as shown here, has recently been disputed (Erusalimsky and Skene, 2009).…”

Endothelial progenitor cells: Implications for cardiovascular disease

“…Is there a possibility to tune EPCs in culture, so that their angiogenic potential is increased? In a recent study Hu et al investigated the effect of low-dose aspirin on the proliferation and the functional capacity of EPCs in cell culture (34). These experiments clearly demonstrated that aspirin up to a concentration of 10-100 lmol/L increased the migratory capacity of the cells without influencing the proliferation.…”

Endothelial Progenitor Cells in Long-Standing Asymptomatic Type 1 Diabetic Patients with or without Diabetic Nephropathy

“…AcLDL uptake and lectin binding have been used in a number of important EPC studies [13,14,15,16,17,18], and constitutes the necessary step in measuring culture EPC number [20]. Additional EPC markers are now recommended.…”

“…MNC (10 × 10 6 ) were seeded in 6-well plates coated with human fibronectin (BD BioCoat, Biosciences) in 3 ml of endothelial basal medium (Lonza) with supplements (EGM SingleQuots: 10 µg/ml hEGF, 1 µg/ml hydrocortisone, 3 µg/ml bovine brain extract, 30 µg/ml gentamicin, 50 µg/ml amphotericin B and 2% fetal bovine serum) as described previously [13,14,15,16,17,18]. After 4 days of culture, nonadherent cells were removed, and adherent cells were washed with phosphate buffer saline before adding 3 ml of fresh endothelial basal medium with supplements, and cultured for 3 days.…”