Effects Of Advanced Glycation End Products On The Cultured Retinal Muller Cells

【Objective】To observe the effects of advanced glycation end products(AGEs) on the cultured retinal Müller cells, in order to further investigate the pathogenesis of diabetic retinopathy (DR).【Methods】In vitro, we cultured human and rabbit retinal Müller cells by way of mixed enzyme digestion and tissue adherence, using human retina of an abortus of 6 months and healthy rabbit. AGEs-BSA and its control were prepared by incubating bovine serum albumin (BSA) with glucose for 12 weeks. Passage Müller cells were divided into AGEs-BSA experimental group, AGEs-BSA control group and blank control group, each group were then be divided into 5 subgroups (8,16,32,64,128μL /200μL) respectively according to concentrations of AGEs-BSA and its control. Cells were treated by AGEs-BSA and its control for 3d, 6d and 9d. Then optical microscope was used to observe the morphological changes of cells. MTT assay was performed to examine the proliferative ability of cells of each group. And flow cytometry assy (FCM) was used to detect cell apoptosis at highest concentration of AGEs-BSA (128μL /200μL) for 3d and 9d.【Results】Cultured human retinal Müller cells manifest typical cell appearance : long fusiform cell body, adequate endochylema and elliptical nucleus in the center of cell body. Under optical microscope, no obvious morphological changes of cells were seen after they were treated by AGEs-BSA for only 3d, while with the extension of time, typical cell appearance could not be seen. By MTT assay, when cells were treated by AGEs-BSA for 3d, the OD values of cells of each group have no significant difference (P>0.05). While after 6 days, the OD values of cells were significant different with those of control groups; after 9 days, the OD values of cells were significant different with those of control groups either. Results of FCM confirmed the apoptosis rates of cells exposed to AGEs-BSA (128μL /200μL) for 3d and 9d were higher than those exposed to its control. Furthermore, the apoptosis rates of cells were positive correlation with interference time of AGEs-BSA.【Conclusion】AGEs-BSA can inhibit the growth of retinal Müller cells, which is related with the concentration of AGEs-BSA and the interference time . This kind of inhibition of AGEs on retinal Müller cells will help us further understand the pathogenesis of DR and find new ways to prevent and treat DR.