Nutr Res Pract.  2014 Aug;8(4):368-376.

Effects of lycopene on number and function of human peripheral blood endothelial progenitor cells cultivated with high glucose

• Zeng YC
• Mu GP
• Huang SF
• Zeng XH
• Cheng H
• Li ZX

Abstract

BACKGROUND/OBJECTIVES
The objectives of this study were to investigate the effects of lycopene on the migration, adhesion, tube formation capacity, and p38 mitogen-activated protein kinase (p38 MAPK) activity of endothelial progenitor cells (EPCs) cultivated with high glucose (HG) and as well as explore the mechanism behind the protective effects of lycopene on peripheral blood EPCs.
MATERIALS/METHODS
Mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation. EPCs were identified after induction of cellular differentiation. Third generation EPCs were incubated with HG (33 mmol/L) or 10, 30, and 50 microg/mL of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation and apoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performed by replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity was assayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylated and non-phosphorylated p38 MAPK.
RESULTS
The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and 50 microg/mL of lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantly lower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirm the causal relationship between MAPK inhibition and the protective effects of lycopene against HG-induced cellular injury, we treated cells with SB203580, a phosphorylation inhibitor. The inhibitor significantly inhibited HG-induced EPC injury.
CONCLUSIONS
Lycopene promotes proliferation, migration, adhesion, and in vitro vasculogenesis capacity as well as reduces apoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-induced EPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-induced EPC injury by inhibiting p38 MAPKs.

Keyword

Lycopene; high glucose; endothelial progenitor cells; p38 MAPK; curve fitting

MeSH Terms

Apoptosis
Blotting, Western
Centrifugation, Density Gradient
Ficoll
Flow Cytometry
Glucose*
Humans
p38 Mitogen-Activated Protein Kinases
Phosphorylation
Protein Kinases
Signal Transduction
Stem Cells*
Ficoll
Glucose
Protein Kinases
p38 Mitogen-Activated Protein Kinases

Figure

• 

  Fig. 1 Characterization of EPCs under laser-scaning confocal microscope (×200). Under the laser confocal microscope, Dil-Ac-LDL (red, excitation wavelength 543 nm) and Ulex lectin FITC-I(green, excitation wavelength 477 nm) double stained positive cells were differentiating EPCs

• 

  Fig. 2 Effects of lycopene on vasculogenesis activity of EPCs (×100). Compared with the other dose lycopene groups(10,50 µg/mL )and signal blockers, microvascular structures of 30 µg/mL lycopene group were more complex and complete. Representative pictures of the tube formation of EPCs with different treatments are shown. NC: normal control group;HG:High glucose group; SB203580:p38 MAPK inhibitor.

• 

  Fig. 3 Effects of lycopene on high glucose induced activation of p38 MAPKs in EPCs. Compare with NC, *P < 0.05;Compare with HG, ▵P < 0.05; Compare with 30 µg/mL lycopene group, ▴P < 0.05. NC: normal control group; HG: High glucose group; SB203580: p38 MAPK inhibitor.

• 

  Fig. 4 Curve fitting figures of effects of lycopene on EPCs in high concentration of glucose

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