| Objective To observe the effect of high glucose on the expression of erythropoietin(EPO) mRNA and protein in human umbilical vein endothelial cells(HUVECs) cultured in vitro and lay a foundation for the study of pathogenesis of diabetic retinopathy(DR).Methods(1)Cultured HUVECs in vitro were divided into two groups:①High glucose group:HUVECs were cultured with high glucose at a concentration of 22 mmol/L for 6h,12h,24h,48h,72h respectively.②Control group:control group 1 is normal control group(5.5mmol/L glucose),control group 2 is mannitol control group(5.5mmol/L glucose+16.5mmol/L mannitol).(2)The expression of EPO mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR).(3)The expression of EPO protein was detected by immuocytochemistry method.Results(1) HUVECs expressed EPO mRNA and protein in high glucose group and control group.(2)Compared to control group 1,the expression of EPO mRNA and protein in HUVECs of control group 2 did not have significant difference(p＞0.05).(3)The expression of EPO mRNA in HUVECs at a high concentration of 22mmol/L glucose for 12h,24h,48h were higher than that in control group 1(p＜0.05).For 72h, the expression of EPO mRNA in HUVECs decreased,which did not have significant difference compared to control group 1(p＞0.05) (4)Immunocytochemistry indicated that EPO protein located in the cytoplasm of HUVECs.The expression of EPO protein in HUVECs at a high concentration of 22mmol/L glucose for 12h,24h,48h were higher than that in control group 1(p＜0.05).For 72h,the expression of EPO protein in HUVECs decreased,which did not have significant difference compared to control group 1(p＞0.05)Conclusion(1) High glucose could increase the expression of EPO mRNA and protein in HUVECs,which is time-related.(2)The effect of high glucose on the expression of EPO mRNA and protein in HUVECs is not related to osmotic pressure.EPO may be an important regulator in neovascularization.
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