| Objective: 1. To investigate the relation between the proliferation, migration and adhering of endothelial progenitor cells(EPCs) in peripheral blood with coronary heart diseases cultured in vitro and the activity of telomerase. 2. To observe the effects of HGF on the proliferation, migration and adhering of endothelial progenitor cells cultured in vitro,and to investigate the relation between the concentration of HGF , the numbers of CD133~+ cells in peripheral blood and the biological function of endothelial progenitor cells.Methods: l.Mononuclearcells(MNCs) were harvested from peripheral blood by density gradient centrifugation,then inoculated in culture plates coated with gelatin. After cultured for 7d, the immunohistochemical method was performed to identify EPCs. The proliferation,migration and adhering of EPCs was measured by MTT assay,transwell insert and the measured experiment of adhering ability. 2. The activity of telomerase was detected by TRAP-PCR argentation. 3. 15 patients with coronary heart disease(CHD)and 15 matched control subjects were enrolled. To observe the change of the number and function of EPCs from patients with CHD. MNCs harvested by density gradient centrifugation were inoculate in culture plate coated with Gelatin. After culture for 4d, wash-out non-adherent cells and attached cells divide into HGF group and non-HGF group, then continue to culture for 3d, to observe the effects of 1ng/μl HGF on the proliferation, migration, adhering and apoptosis of endothelial progenitor cells. 4. MNCs in periheral blood were harvested by density gradient centrifugation, and marked with FITC-Conjugated Mouse Anti-human CD133 then the number of CD133~+ cells were detected by flow cytometry; The concentration of HGF in blood plasma was determined by the method of ELISA. Results: 1. EPCs were well isolated from human peripheral blood.The proliferation (P<0.01),adhering (P<0.01) and migration (P<0.05) of EPCs isolated from patients with CHD were reduced, compared with the control group. 2.The activity of telomerase were reduced in patients with CHD, compared with the control group (P<0.05) . 3.1ng/μl HGF can significantly improve the ability of proliferating (P <0.05), adhering (P<0.01) and migrating (P<0.05) of EPCs isolated from patients with CHD and improve the ability of proliferating (P<0.05) with non-CHD, but had not this effect of adhering and migrating in the control subjects (P>0.05) . EPCs apoptosis was not significantly different between CHD and non-CHD (P>0.05) . HGF had not a protecting effect on EPCs apoptosis (P>0.05 ) .2. Compared with patients with non-CHD,the number of CD133~+ cells in periheral blood of patient with CHD was lower (P<0.01) and the concentration of HGF in blood plasma was higher (P<0.01) .Conclusions: 1. The proliferation,adhering and migration activities of EPCs are decreased in patients with CHD. The weakening of EPCs biological function might be correlated with the attenuation of telomerase activity. 2. HGF could improve all biological functions of EPCs in patients with CHD and improve the ability of proliferating with non-CHD.but had not this effect of adhering and migrating in patients with non-CHD . 3. EPCs apoptosis was not significantly different between CHD and non-CHD. HGF had not a protecting effect on EPCs apoptosis. 4. It was promising auxiliary diagnosis and the way of examine for CHD to detect the number of CD133~+ cells and the concentration of HGF in periheral blood.
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