Effects of lycopene on number and function of human peripheral blood endothelial progenitor cells cultivated with high glucose
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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 peripheralblood 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 μg/mL of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation andapoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performedby replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity wasassayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylatedand non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and 50 μg/mLof lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantlylower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirmthe 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 reducesapoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-inducedEPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-inducedEPC injury by inhibiting p38 MAPKs.
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 peripheralblood 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 μg/mL of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation andapoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performedby replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity wasassayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylatedand non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and 50 μg/mLof lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantlylower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirmthe 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 reducesapoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-inducedEPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-inducedEPC injury by inhibiting p38 MAPKs.
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