Effects Of Icariine On The Numbers And The Expression Of Nitric Oxide (NO) Secretion Of Endothelial Progenitor Cells Cultured In Vitro From Peripheral Blood

Objective:To explore the effects of icariine on the numbers and the expression of NO secretion of endothelial progenitor cells(EPCs) cultured in vitro from peripheral blood.Methods:(1).EPCs were isolated by density gradient centrifugation from human peripheral blood,cultured with Medium 199. (2). EPCs were characterized as adherent cells double positive for Dil-acLDL uptaking and FITC-lectin binding by laser confocal microscope,and further demonstrated by expressing CD34 and CD 133 using flow cytometry. (3).EPCs inverted microscope observation of the morphological changes. (4).Cultured EPCs were rando-mized into 3groups:control group, atorvastatin group(10μmol/L) and icariine group(10,25,50ng/ml). (5). Numbers of EPCs was assessed by cell counting and NO secretion of EPCs was assessed.Results1.Characteristics of human EPCsAfter 7 days culture the MNCs showed as spindles-shaped,endothelial cells-like morphology and colony-like growth into cell fusion. Fluorescence microscopy revealed absorption Dil-acLDL and FITC-UEA-1,Dil-acLDL uptake in the cells were red,cells lectin binding were green,double positive cells appearing yellow. Further the cells were demonstrated by expressing CD34+(38.36%±3.7%) and CD133+(13.91%±1.47%) using flow cytometry.2.Effects of icariine on the numbers and the expression of nitric oxide (NO)secretion of EPCsThe number of endothelial progenitor cells were remarkably increased both in atorvastatin group and different concentration ICA groups,which was most prominent at the concentration of ICA of 50ng/ml for 48 hours. Compared with control group,25 and 50ng/ml icariine group showed higher NO production and 48 hour group is better than 24 hour group in function; atorvastatin group showed the highest NO production.Conclusion1.Through the method of Ficoll density gradient centrifugation can be isolated and cultured EPCs from human peripheral blood.2. ICA increase the numbers and improve the ability of NO secretion of endothelial progenitor cells(EPCs). 3. ICA could improv the number and function of EPCs.It's likely to be a new mechanism of icariine (ICA) to prevent and treat vascular endothelial injury/dysfuntion-related disease.