Study On The Biological Characteristics Of Bone Marrow Endothelial Progenitor Cells From Diabetic Mice After Mobilization And Culture

Objective(s):To investigate the changes of characteristics and biological activities of EPCs after mobilization from diabetic high glucose microenvironment.and to provide evidence for diabetic wound healingMethods:AMD3100 was combined with recombinant human granu-locyte colony-stimulating(rhG-CSF)to mobilize endothelial progenitor cells(EPCs).?db/db group:10 db/db male rats were taken,each mouse was injected with AMD3 1 00 for the first time,G-CSF was continuously injected for 5 days,and EPCs were prepared by blood sampling on the 10th day.?db/+group:10 db/+male rats were taken,each mouse was continuously injected with G-CSF for 5 days,AMD3100 was injected once every day,and EPCs were prepared by blood sampling on the 7th day Flow cytometry was used to detect the proportion of CD34+/CD133+/CD309+cells PCR and Western blot were used to detect Vascular endothelial growth factor(VEGF)and Stromal cell-derived factor(SDF-1).Chemokine receptor 4(CXCR4)gene and protein expression.Results:? CCK-8 experiments showed that at the same time point,the proliferation rate of db/+group was significantly higher than that of db/db group;at different time points,the proliferation rate increased gradually with time(p=0.000);?No significant difference in adhesion function between the two groups(p>0.05):two groups There were no significant differences in migration(p>0.05)and lumen formation(p>0.05).?The expression levels of VEGF,CXCR4 and SDF-1 in db/db group were lower than those in db/+group(p=0.019;p=0.001;p=0.003);?The mRNA expression levels of VEGF,CXCR4 and SDF-1 in db/db group were lower than those in db/+group(p=0.029;p=0.009;p=0.009).Conclusion(s):The method of mobilizing diabetic bone marrow endothelial progenitor cells with AMD3100 and G-CSF can isolate more high-purity EPCs from peripheral blood The proliferation,migration and tube formation functions of EPCs in diabetic state are impaired.Although the expression level of related angiogenic factors in cultured endothelial progenitor cells has improved,it is still lower than that of normal EPCs.