Changes In The Number And Function Of Endothelial Progenitor Cell From Peripheral Blood In Patients With Diabetic Retinopathy

ObjectiveVascular endothelial dysfunction is one initiating factor of happening and developing about diabetic retinopathy(DR). Endothelial progenitor cell (EPC) is such a cell which can participate in vascular regeneration after birth and repair vascular endothelial injury. The purpose of our research is to investigate the changes in the number and activity of endothelial progenitor cell from peripheral blood in patients with diabetic retinopathy. Discussing the role of EPC in the pathogenesis of diabetic retinopathy may provide us theoretical basis to prevent and control early diabetic retinopathy, and also give us a reference for clinical treatment.MethodsA case-control study included:12patients with DR,18patients with unique DM without any complications of DR and15patients without DM as control. Peripheral blood mononuclear cells were isolated by density gradient centrifugation. After10days induced differentiation, identified the adherent cells were EPC by fluorescein staining and flow cytometry. Proliferation, adhesion and migration activities of EPC were assayed by MTT chromatometry, adhesion activity assay essay and Transwell assay.ResultsMNCs shaped round or oval were isolated by Ficoll density gradient centrifugation. Trypan blue staining measured the viabilities of MNCs were over99%. In the morning of culture, most cells became larger, and a part of round MNCs attached dishes become fusiform EPC after3-5days. About in the seventh day, cell colonies contains several cells emerged. Cells become long-spindle form in the tenth day. After15-20days, cells assumed crazy paving appearance. The function of EPC for Dil-ac-LDL uptaking and FITC-UEA-1binding was both strong positive in tenth day. Flow cytometric test showed that positive rate of EPC in patients with DR group, DM group and control group was (37.375±2.501)%,(30.131±3.245)%and (45.192±1.287)%, there was significant difference of3groups (F=27.690, P=0.001). MTT chromatometry showed that OD)-value of DR group, DM group and control group was0.330±0.047,0.225±0.042and0.120±0.029, there was significant difference of3groups(F=29.327, P=0.000). Adhesion activity assay essay showed that EPC numbers in DR group, DM group and control group was76.400±7.503,51.167±6.646and26.500±7.853, there was significant difference of3groups(F=56.612, P=0.000). Transwell assay showed that EPC numbers in DR group, DM group and control group was23.600±6.504,20.833±4.491and12.000±2.944, there was significant difference of3groups (F=6.477, P=0.012).Conclusions1. Endothelial progenitor cells from peripheral blood in patients with diabetic retinopathy exhibit reduced numbers and impaired proliferation, adhesion and migration activity.2. Reduced number and impaired function of peripheral blood EPC may be the key link of diabetic retinopathy occurrence and development.3. Research of peripheral blood EPC number and function may become a new target for prevention and treatment of diabetic retinopathy.