PEGFP/Ang-1 Transfected BMSCs Attenuates High Glucose Concentration-induced Choroido-retinal VECs Damage

ObjectiveTo investigate the beneficial effects of recombinant plasmid (pEGFP/Ang-1) transfected rats bone marrow mesenchymal stem cells (BMSCs) on retinal endothelial cells (RF/6A).MethodsUsing adherence screening and density gradient centrifugation methods, we obtained purified rats bone marrow stem cells (BMSCs). After transfection with recombinant plasmid (pEGFP/Ang-1), the BMSCs were screened with G418, and the positive clones were then sequentially confirmed by expression of enhanced green fluorescent protein (GFP) with fluorescence microscope, and expression of angiogenin-1 (Ang-1) using Western blot method. To explore the beneficial effects of pEGFP/Ang-1 transfected BMSCs on RF/6A under high glucose concentration, the transfected BMSCs were cocultured with rhesus chorioido-retinal endothelial cells (RF/6A) in transwell under high glucose concentration. Furthermore, the vitality, DNA ladder and pPKB of RF/6A were detected with methyl thiazolyl tetrazolium (MTT) motheds, agarose gel electrophoresis (AGE) and Western blot respectively.ResultsThe successfully stable transfection of pEGFP-N1-Ang-1 in BMSCs, can be stably expressed Ang-13 days.later started to diminish. Transwell co-culture with RF/6A in d 3RF/6A inferior chamber for testing. MTT colorimetric assay detect the cell is survival, it can detect cell survival that with the prolongation of glucose, the greater the damage to the cells. Plasmid pEGFP-N1-Ang-1 protein in cells has a protective effect on RF/6A. DNA agarose electrophoresis shows, B cells are not detected DNA ladder, the other groups have different levels of expression of DNA ladder; western blot detection of phosphorylated protein kinase B expression, showing that increased expression of PKB, indicating that on cells inhibit apoptosis.Conclusion1 High concentrations of glucose can lead to produce RF/6A toxicity. Restructuring plasmid carrying pEGFP/Ang-1 BMSCs can protect the high concentration of glucose miniaturization endothelial cell.2 After BMSCs can express the carrying Ang-1, increase, and preventPKB phosphorylation of RF/6A high concentration of glucose the damage. Keywords vascular eery-1; Mesenchymal stem cells; Choroidal-retinal endothelial cells...