[Effects of puerarin on number and activity of endothelial progenitor cells from peripheral blood].
Zhang Fr, Chen Jz, Zhu Jh, Wang Xx, Jian-Hua Zhu, Yun-Peng Shang, Xiao-Gang Guo, He-Min Dai, Jian Sun
Abstract
Zhang Fr, Chen Jz, Zhu Jh, Wang Xx, Jian-Hua Zhu, Yun-Peng Shang, Xiao-Gang Guo, He-Min Dai, Jian Sun
Abstract
OBJECTIVE: To investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferation, migration and adhesion. METHOD: Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days culture, attached cells were stimulated with puerarin (to make a series of final concentrations: 0. 1, 0.5, 1, 3 mmol x L(-1)) or vehicle control for the respective time points (6, 12, 24, 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, then adherent cells were counted. RESULT: Incubation of isolated human MNCs with puerarin dose increased the number of EPCs, maximum at 3 mmol x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, puerarin also promoted EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity. CONCLUSION: Puerarin can augment the number of EPCs with enhanced functional activity.
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OBJECTIVE: To investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferation, migration and adhesion. METHOD: Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days culture, attached cells were stimulated with puerarin (to make a series of final concentrations: 0. 1, 0.5, 1, 3 mmol x L(-1)) or vehicle control for the respective time points (6, 12, 24, 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, then adherent cells were counted. RESULT: Incubation of isolated human MNCs with puerarin dose increased the number of EPCs, maximum at 3 mmol x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, puerarin also promoted EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity. CONCLUSION: Puerarin can augment the number of EPCs with enhanced functional activity.
Key concepts: Puerarin, Vasculogenesis, Ficoll, Progenitor cell, In vitro, Peripheral blood mononuclear cell, Molecular biology, Andrology