Promoted Endothelial Progenitor Cells Proliferation Research Articles

Cyclosporin A Suppresses Proliferation of Endothelial Progenitor Cells: Involvement of Nitric Oxide Synthase Inhibition

To investigate the effects of the potent immunosuppressive agent cyclosporin A (CsA) on the proliferation of human endothelial progenitor cells (EPCs) and endothelial nitric oxide synthase (eNOS) expression in EPCs. The EPCs were obtained from cultured mononuclear cells, which were isolated from the peripheral blood of healthy adults, and stimulated with CsA (10 microg/mL) in the presence or absence of either vascular endothelial growth factor (VEGF; 50 ng/mL) or L-arginine (1 mM). To explore the effect of different concentrations of CsA alone on EPC proliferation, some cells were treated with CsA in a series of final concentrations ranging from 0 to 10 microg/mL. Cell proliferation and apoptosis were determined, respectively, by the Cell Counting Kit-8 assay and terminal deoxynucleotidyl transferase-mediated nick end labeling staining. The expression of eNOS was assayed by reverse transcription-polymerase chain reaction analysis while nitric oxide (NO) generation was detected using the Griess method. The effects of CsA on EPC proliferation, apoptosis, and eNOS/NO production were dose dependent in the concentration ranging from 0.1 microg/mL to 10 microg/mL. Treatment with VEGF (50 ng/mL) significantly promoted EPC proliferation and eNOS/NO production, which were completely abrogated by pre-incubation with CsA (10 microg/mL). The supplement of L-arginine (1 mM) promoted NO production that enhanced EPC proliferation and attenuated the effect of CsA on EPC proliferation and apoptosis. CsA significantly inhibited proliferation, eNOS mRNA expression and NO production of human EPCs, in a dose-dependent manner.

EFFECTS OF RESVERATROL ON NUMBER AND ACTIVITY OF ENDOTHELIAL PROGENITOR CELLS FROM HUMAN PERIPHERAL BLOOD

1. It has been well established that oestrogens can increase the number of endothelial progenitor cells (EPC) by anti-apoptotic effects. Resveratrol, a polyphenolic phytoalexin extracted from grapes and wine, has been reported to act as an oestrogen receptor agonist. We hypothesize that putative phyto-oestrogen may promote EPC proliferation and survival in vitro. 2. Endothelial progenitor cells were isolated from human peripheral blood and identified immunocytochemically. Endothelial progenitor cells were incubated with resveratrol (1, 10, 25 and 50 mmol/L) or control for specified times. Cell proliferation, migration and in vitro vasculogenesis were assayed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra-zolium bromide (MTT) assay, modified Boyden chamber assay and in vitro vasculogenesis detection, respectively. 3. Resveratrol increased the number of EPC and promoted EPC proliferation, adhesion and migration in a dose- and time-dependent manner. Cell number peaked at 50 mmol/L resveratrol after incubation for 24 h compared with vehicle control (61.3 +/- 5.8 vs 112.8 +/- 7.2, respectively; P < 0.01). 4. Furthermore, cell cycle analysis showed that 50 mmol/L resveratrol significantly increased the S phase and decreased the G(0)/G(1) phase of EPC. In addition, resveratrol increased vascular endothelial growth factor production and further induced vasculogenesis in vitro. 5. In conclusion, resveratrol significantly induces EPC proliferation, migration and further promotes angiogenesis in vitro.