Protection Effect Of RhEPO To Apoptosis Of Retinal Nerve Cells In High Glucose And Expression Of Bcl-2 In Vitro

Objective To investigate the protective effects of Recombinant Human Erythropoietin (rhEPO) to apoptosis of retinal on apoptosis retinal nerve cells mixed cultured in high glucose in vitro and the expression of Bcl-2.Methods A stable and reliable method to culture retinal nerve cells by trypsin(0.125%) digstion dissecting the retinaes of rats postnatal 1-3 days in vitro. Trypan blue staining was used to observe the modality, grewing condition and survival status of nerve cells in different concentrations of glucose. Immunocytochemical technique was used to indentify the retinal neurons, such as anti-rat neuron-specific enolase(NSE) antibody. The primary retinal cells of postnatal rats in 1-3 days had been cultured for 3 days in vitro and were randomly divided into high glucose group, control group, and therapy group by different concentrations of rhEPO. The retinal cells in the treatment group have been cultured for 48h with high glucose (5,10,20,40U/ml), The method of MTT was used to estimate the vitality of retinal cells, and the expression level of Bcl-2 of nerve cells was detected by semi-quantitative immunocytochemistryResults The results of immunocytochemical staining demonstrate that most of the cells were positive in vitro for anti-NSE antibody responses and the purity of retianl cells were above 90%. At the condition of high glucose, the vitality of retinal cells decreased and many cells were apoptoic by interfering with 30 mmol/L glucose for 48 hours. The result showed that rhEPO had a protective effect on retinal cells injuried by high glucose.All the group of rhEPO could effectively reinforce the expression of Bcl-2 protein, and the protective effect was positively related with the concentration and time.Conclusion rhEPO could offer the protective effect to the apoptosis of neuronal cells injuried by high glucose in vitro, and the neuroprotective effect has the time dependency and concentration dependency. The result shows that the high expression of Bcl-2 may be one of the mechanisims of its protection to apoptosis of neuronal cells.