| Purpose and Background:Endothelial progenitor cells(EPCs) are precursor cell of endothelial cells.EPCs have characteristics of self-proliferation，homing directionally and can bedirected to differentiate into mature endothelial cells. Prospects of clinicalapplication are broad. Recent studies have provided a favorable evidence thatneovascularization after the birth don’t only rely on preexisting vascularsprouting, but also on the EPCs from bone marrow and circulating blood.Onlyif endothelial integrity,vascular normal function structure can keep.It isreported that the reduction of EPCs had related to cardiovascular disease riskfactors. Volume and quality of EPCs from the circulating blood are and closelyrelated to repair endometrial. EPCs involved in the vascular endothelial cellsrepair process.One of the purposes of this study is to determine the isolation and cultureof endothelial progenitor cells by VEGF165and bFGF from peripheral blood inhealthy adults,and reseach cell shape,amount,activity and draw growth curve.The second purpose of this study is to investigate the isolation and vultureof endothelial progenitor cells from peripheral blood in patients with SAP,ACSand normal people.And study the correlation of EPCs number and the degree ofcoronary artery stenosis and risk factors aggregation.As a first-line drug for the prevention and treatment of coronary heartdiseases, HMG-CoA reductase inhibitors-statins on lipid-lowering cholesteroldrugs have been confirmed with a number of cardiovascular protection,including the improvement of endothelial function, inhibiting of smooth musclecell proliferation and migration,the inhibition of platelet aggregation, and theinflammatory response of microvascular. Some recent studies have showed that statins also promote angiogenesis after acute ischemic, accelerate endothe-lialization of vascular injury induced by balloon and reduce neointimalthickening, but the two roles are closely related with the EPCs.The third purpose of this study is to observe the effects on number、expand、adhesion and migration ability of EPCs of patients with SAP indifferent concentrations of rosuvastatin.Materials and Methods:Experiment one：Mononuclear cells isolated from peripheral isolatedblood of healthy adults by Ficoll-density centrifugation.The cells werecultured in1640medium supplemented with VEGF165and bFGF.The EPCspecific surface mark CD34、CD133and KDR were assessed by fluorescenceactivated cell sorter (FACS) analysis. After in vitro cultivation, diferentiatingEPCs were identified asadherent cells double positive for Dil-acLDL andFITC-UEA-1by direct fluorescentstaining under a laser scanning confocalmicroscope, migration was assayed by MTT assay, and reseach cell shape,amount, activity and draw growth curve.Experiment two：From June2008to May2010, patients diagnosed ascoronary heart disease were enrolled and divided into three groups: acutecoronary disease（ACS group，n=70），stable angina group (SAP group, n=56),patients with normal coronary angiography were enrolled as control (controlgroup, n=49). EPCs proliferation and migration activity were detected withMTT assay and modified Boyden chamber assay,respectively. EPCs adhesionassay was performed by replating MNCs on the humanfibronectin coatedplates,and the adherent cells were counted. Analysis the change on number、expand、adhesion and migration ability of EPCs of patients with ACS, SAPand normal coronary. And study the correlation of EPCs number and thedegree of coronary artery stenosis and risk factors aggregation. Experiment three：EPCs from stable angina group were cultured indifferent concentrations of rosuvastatin, observe the change on number、expand、adhesion and migration ability.Statistics：Numerical data are expressed as mean±standard deviation (M±SD).SPSS13.0was used to analyze the data. Anova were used to compare amongmultiple groups, and q test for between groups. t test for comparing means oftwo groups. p<0.05was considering as statistical difference.Results：Experiment one：Mononuclear cells isolated from peripheral blood will beinduced endothelial progenitors cells by VEGF165and bFGF. Endothelialprogenitors cells assessed by fluorescence activated cell sorter (FAGS)analysis、 laser scanning confocal microscope. Epc have good migrationcapacity by VEGF165and bFGF induced.Experiment two：The number of EPCs was significantly reduced inpatients with SAP compared with control subjects, and EPC function (expand、adhesion and migration ability)were decresed. In addition, EPCs number andfunction were decresed accompaniment with the degree of coronary arterystenosis. The number of EPCs was increased remarkably in patients with ACScompared with control subjects, but EPCs function (expand、adhesion andmigration ability)were decresed, and these change become heavieraccompaniment with the degree of coronary artery stenosis and have nocorrelation with the extend of coronary artery stenosis. In many risk factors thatcan induced coronary heart disease, the number of EPCs have significantcorrelation with aging, hypertension, smoking, high-LDL.The amount of EPCswas reduced accompanied by the increased risk factors number. The two partshave negative correlation. The number of EPCs and inflammatory factor C respone proteim increased remarkably in patients with ACS, and the two partshave positive correlation. Gensini score and the level of C respone proteimincrease gradual accompanied by the degree of coronary artery stenosis.Gensini score and the level of C respone proteim have positive correlation. Thelevel of C respone proteim in SAP and control group was similar(P＞0.05).Experiment three： HMG-CoA reductase inhibitors can significantlyincrease the number and expand、adhesion and migration ability of EPCs. Thereis a dose-dependent effect between rosuvastatin, the changes become apparentalong with gradual increase concentrations of rosuvastatin., and reach peak atthe concentration of1umol/L. In the same concentration of rosuvastatin thenumber and expand、adhesion and migration ability of EPCs have no releventto the length of time of culturing..Conclusion:Experiment one：Mononuclear cells isolated from peripheral blood will beinduced endothelial progenitor cells under certain conditions.Experiment two：The number of EPCs was significantly reduced inpatients with SAP compared with control subjects, and EPCs function (expand、adhesion and migration ability)were decreased. The number of EPCs wasincreased remarkably in patients with ACS compared with control subjects, andEPCs function (expand、adhesion and migration ability)were decreased, too,andthese change become heavier accompaniment with the degree of coronaryartery stenosis. The number of EPCs have significant correlation with aging,hypertension, smoking, high-LDL. The amount of EPCs was reducedaccompanied by the increased risk factors number. The two parts have negativecorrelation.Experiment three： HMG-CoA reductase inhibitors can significantlyincrease the number and expand、adhesion and migration ability of EPCs, and reach peak at the certain concentration. The number of EPCs and inflammatoryfactor C respone proteim increased remarkably in patients with ACS, and thetwo parts have apparent positive correlation. |
PDF Full Text Request