| objective To investigate the effect of recombinant human erythropoietin (rhEPO) on glutamamine synthetase (GS) rat retinal mUller cells induced by high glucose in vitro and the possible mechanism.Methods Retina of postnatal1-3day SD rats was dissected into cell suspension by using trypsin digestion. Trypan blue staining was used to study the effcts of different concentrations of glucose on the morphology, growth and survival status of retinal neurons.The retinal mUller cells of postnatal SD rats were randomly divided into control group,high glucose group,and treatment group with different concentrations of rhEPO.The retinal mUller cells in treatment group were cultured in high glucose medium with510、20、40U/ml rhEPO. After48hours, the apoptosis of retinal mUller cells were assayed by using TUNEL, and the expression levels of GS protein of mUller cells were detected with semi-quantitative immunocytochemistry.Results Immunocytochemical staining showed that most of the cells in vitro positive for anti-GS antibody responses, the purity of retinal mUller cell was above90%. The vitality of retinal neurons in high glucose decreased and a lot of cells were apoptoic, interfering with mUller cells with25mmol/L glucose for 48hours was as the high glucose model. Application of exogenous rhEPO had a protective effect on with injury ofmuller cells high glucose, adding540U/ml rhEPO treatment could effectively promotoed the expression of caspase-3protein (P<0.01), and which was positively related to concentration.Conclusion In mimicked diabetic condition, rhEPO treatment can protect mUller cells from apoptosis induced by high glucose injury, and its neuroprotective effect is related to concentration. Inhibiting the expression of GS protein may be one of the mechanisims of its protection,... |
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