The Effects Of Recombinant Human Erythropoietin On Apoptosis Of Rat Muller Cells Cultured In High Glucose And The Expression Of Clusterin

Objective To explore the effects of Recombinant human erythropoietin(rhEPO)on apoptosis of rat retinal Miiller cells cultured in high glucose in vitro and the expression of Clusterin and to probe the mechanism.Methods The retinas of Sprague-Dawley(SD)rats at postnatal 3 days were isolated,chopped into small fragments and cultured in vitro by means of enzyme digestion.Muller cells were purified with mechanical shaking when the medium was replaced.The cells we cultured were indentified by immunocytochemical staining,hematoxylin-eosin staining and TEM.We divided the Purified retinal Miiller cells into control group,high glucose group,and therapy group by rhEPO randomly.The retinal cells were cultured for 48h in the treatment group with high glucose of 5 kU/L、10 kU/L、20 kU/L、40kU/L rhEPO.The vitality of retinal Muller cells were measured by The method of MTT.We took enzyme-linked immuno sorbent assay(ELISA)to test and the expression levels of CLU Both inside and outside the retinal Miiller cells.Results The purified Mullers cells with hematoxylin-eosin staining had irregular shape,abundant cytoplasm,2 or more 2 elliptic nucleus were long and narrow.The TEM showed that purified Mullers cells with many microvillus on the surface were had Abundant organelles and contained a large number of characteristic microfilament of 8-10nm.The results of GS immunocytochemical staining certified that over 90%of the purified Mullers cells were positive.At the high glucose group,the vitality of retinal Muller cells dcreased contrast to the control group,the expression of CLU protein outside the cell dcreased and the expression of CLU protein in the cell increased(P<0.05).The expression of CLU protein of the groups of rhEPO intervention increased effectively.while the expression of CLU protein outside the cells decreased(P<0.05).Conclusion The apoptosis of rat retinal Muller cells in high glucose could be restrained by rhEPO in vitro.Its mechanism may be that the sCLU that resist apoptosis rise,the nCLU that promote apoptosis decline,Then,the mechanism will regulate sCLU/nCLU,which lead to inhibition of apoptosis,and protect the cells from apoptosis eventually.