The Effect Of Rosuvastatin On Proliferation And Apoptosis Of Endothelial Progenitor Cells From Peripheral Blood In CHD Adults

Objective：To investigate the effect and the mechanism of Rosuvastatin on the proliferationand apoptosis of EPCs from peripheral blood in patients with coronary heart diseases(CHD).Methods：Thirty six patients in the department of cardiology of the third affiliated hospitalof nanchang university with angiographically documented CHD were enrolled in thepresent study from October2011to October2012. Under the aseptic conditions,20mlheparin anticoagulation peripheral blood from the Radial artery or Femoral artery wascollected and processed within4hours. The patients who have the recent trauma,surgery, skin ulcers, inflammation, tumor or blood system diseases were excluded.Mononuclear cells (MNCs) were isolated by density gradient centrifugation, then thecells were plated on fibronectin-coated culture dishes.After being cultured for7days,the attached cells were divided into4groups:1.control group: the attachedcells were incubated in RPMI1640medium without FBS for the first24hours, thenthey were incubated in the medium supplemented with FBS for24hours;2.Rosuvastatin groups (the last groups): after being incubated in the medium withoutFBS for24hours, the attached cells were incubated with different concentrations ofRosuvastatin respectively (0.01,0.100,1.00umol/L) for24hours. EPCs weredocumented by a laser scanning confocal microscope, EPCs adhesion assay wasperformed by replating it on fibronectin-coated dishes, and then the adherent cellswere counted. EPCs proliferation, migration were detected by MTT assay andmodified Boyden chamber assay respectively. Flow cytometry was used to detect theEPCs apoptosis rate. The expressions of bcl-2mRNA and protein were detected byRT-PCR and Western blot respectively.Result:1.The identification of EPCs After being cultured7days, MNCs isolated from the patients with CHD formedspindle sample of endotielial cells, EPCs were characterized as adherent cells doublepositive for Di-acLDL uptake and lectin binding by direct fuorescent stainning underalaser scanning confocal microscope. EPCs were further documented by theexpression of Bio-markers CD34, CD133and VEGFR-2by flow cytometry.2. The effect of Rosuvastatin on the proliferation and apoptosis of EPCsRosuvastatin remarkably promoted EPCs proliferation, adhesion and migrationand decreased the apoptosis rate in a concentration-dependent manner.3. The effect of Rosuvastatin on the expressions of bcl-2mRNA and protein ofEPCsIn normal condition, the EPCs express the bcl-2mRNA and protein. Theexpressions of mRNA and protein of bcl-2were promoted in EPCs by Rosuvastatinin a dose-dependent manner.Conclusion:1. The Rosuvastatin promotes the proliferation, adhesion and migration of theEPCs in vitro in a concentration-dependent manner;2. The Rosuvastatin can inhibit the apoptosis of the EPCs in vitro by up-regulating the bcl-2expression in a dose-dependent manner.