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Objective: Isolation, culture, human peripheral blood progenitor cells, the effects of different concentrations of icariin and atorvastatin on endothelial progenitor cells (EPCs) and NO secretion of quantity. Method: 1, density gradient centrifugation cultured human peripheral blood EPCS. 2, immunofluorescence staining cells absorb Dil-acLDL and FITC-UEA-I case, flow cytometry detection of EPCs surface markers CD133, CD34 antigen expression. 3, EPCs were observed under inverted microscope morphological changes. 4 experimental groups: the adherent cells were randomly divided into control group, icariin group (final concentration of 10,25,50 ng / ml), atorvastatin group (final concentration of 10μmol / L). 5, using cell counts were observed changes in the number of EPCs, nitrate reductase assay content of NO in the cell culture medium in order to understand its impact on the secretory function of EPCs. Results 1 Dynamic observation under an inverted microscope visible: the first two days in culture cell clusters began to appear the phenomenon; first four days beginning from round into fusiform cells adherent cells; cells cultured on the 7th, the majority of spindle-shaped cells or polygonal, and there colony-like structure appears. 2, can be seen under fluorescence microscopy absorb Dil-acLDL, FITC-UEA-I cells were the red, green fluorescence, two double yellow fluorescent dye is considered differentiating EPCs; flow cytometry shows adherent cells CD34-positive (38.36 ± 3.7)? 133 positive (13.91 ± 1.47)%, further confirming the cultured cells as EPCs. Three different concentrations of icariin group, atorvastatin group and EPCs cultured for 24 h, compared with the control group could display the number of EPCs significantly improved, and the number of EPCs with icariin concentration increases. Icariin 50ng/ml when the most significant improvement in the number of cells of 50ng/ml the aging effects of icariin study showed that all groups showed a time-dependent increase in the number of EPCs, 48h peak. 4, in vitro culture conditions, 10ng/ml icariin group compared with the control group observed indicators EPCs culture supernatant NO concentration difference is not obvious, 25,50 ng / ml compared with the control group EPCs culture supernatant The NO concentration was significantly improved, and 48 hours group than in the 24 hours group effect increased. Atorvastatin group compared with control group, EPCs culture supernatant NO concentration increased significantly. Conclusion 1, using the density gradient centrifugation method can be successfully isolated from human peripheral blood and cultured vascular EPCs. 2, icariin can increase the number of EPCs in human peripheral blood, improve the ability of EPCs NO secretion, and showed a certain amount of time-and-effect relationship. 3, icariin can improve the number and function of EPCs, which may be as a treatment for vascular endothelial injury and endothelial dysfunction-related disease drug mechanisms.
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