| Objective To investigate the protective effects of recombinant human erythropoietin (rhEPO) on the apoptosis of cultured rat retinal neurons induced by high glucose in vitro and the possible mechanism.Methods Retina of postnatal 1-3 day SD rats was dissected into cell suspension by using trypsin (0.125%) digestion. Immuniocytochemistry identified retinal neurons by using anti-rat neuron-specific enolase(NSE) antibody after three days. 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 primary retinal neurons of postnatal SD rats were cultured in vitro for three days and were randomly divided into control group,high glucose group,and treatment group with different concentrations of rhEPO.The retinal neurons in treatment group were cultured in high glucose medium with 5,10,20,40U/ml rhEPO. After 48 hours, the vitality of retinal neurons were assayed by using MTT. The expression levels of caspase-3 protein of retinal neurons were detected with semi-quantitative immunocytochemistry.Results Immunocytochemical staining showed that most of the cells in vitro positive for anti-NSE antibody responses, the purity of retinal neurons was above 90%. The vitality of retinal neurons in high glucose decreased and a lot of cells were apoptoic, interfering with retinal neurons with 30mmol/L glucose for 48 hours was as the high glucose model. Application of exogenous rhEPO had a protective effect on retinal neurons with injury of high glucose, adding 5-40U/ml rhEPO treatment could effectively inhibit the expression of caspase-3 protein (P<0.01), and which was positively related to concentration.Conclusion rhEPO treatment can protect neuronal cells from apoptosis induced by high glucose injury, and its neuroprotective effect is related to concentration. Inhibiting the expression of apoptosis factor caspase-3 may be one of the mechanisims of its protection. |
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