促红细胞生成素对人视网膜色素上皮细胞氧化损伤保护作用及其机制

目的 观察促红细胞生成素（EPO）对人视网膜色素上皮（hRPE）细胞抗过氧化氢（H₂O₂）损伤的影响。方法以传代培养hRPE细胞为研究对象, 采用800μmol/L H₂O₂ 作用3 h 建立 hRPE 细胞损伤模型, 分为正常对照组、单纯损伤组、重组人EPO（rhEPO）治疗组。治疗组又根据加入rhEPO浓度不同分为10、20、40、60 IU/ml亚组。免疫组织化学方法检测细胞中核因子kappa B(NF-κB)的活化情况，比色法测定细胞脂质过氧化产物丙二醛（MDA）含量及乳酸脱氢酶（LDH）细胞释放率。结果 H₂O₂ 可使培养液中MDA及LDH释放率上升，单纯损伤组与正常对照组相比，差异有统计学意义（t_LDH＝29.746，t_MDA＝20.426，P＜0.05）；各治疗组与单纯损伤组比较，MDA及LDH释放率均有显著降低，差异有统计学意义（t_10IU＝5.770，t_20IU＝12.774，t_40IU＝19.818，t_60IU＝24.833，P＜0.05；MDA t_10IU＝5.345，t_20IU＝10.278，t_40IU＝18.571，t_60IU＝20.247，P＜0.05）；各治疗组相关分析显示，rhEPO浓度与LDH细胞释放率及MDA含量呈负相关（r=-0.976，P=0.024； r=-0.968，P=0.032）；rhEPO浓度与NF-κB核移位率呈正相关（r=0.998，P=0.002）；NF-κB核移位率与MDA含量呈负相关（r=-0.954，P=0.046）。结论 EPO能有效地拮抗 H₂O₂ 对hRPE细胞的脂质过氧化损害，其机制可能与活化NF-κB有关。

The protective effects and mechanism of erythropoietin on human retinal pigment epithelial cells during oxidative injury

Objective To investigate the protective effect and mechanism of erythropoietin(EPO)on injury of human retinal pigment epithelial(hRPE)cell induced by hydrogen peroxide(H2O2).Methods Take subcultured hFRPE cells as study target.They were treated with 800μmol/L of H2O2 for3 hours to establish the eell injury model.The cultured cells were divided into three groups:control group,simply injury group and therapeutic group which again divided into 10 IU/ml,20 IU/ml,40 IU/ml,60IU/ml subgroups according to the concentration of recombinant human erythropoietin(rhEPO).NF-kBwas measured by immunohistochemistry.The content of Malondialdehyde(MDA)which was the productof cellular lipid peroxidation and the releasing rate of lactate dehydrogenase(LDH)were estimated bychromatometry. Results H2O2 could elevate the level of MDA and the releasing rate of LDH,comparedsimply injury group with control group,the differences were significant.(tLDH=29.746 tMDA=20.426,P＜0.05);Compared all of therapeutics groups with simply injury group,the releasing rate of MAD and LDHwere decreased obviously,the differences were significant.(LDH t10IU=5.770,t20IU=12.774,t40IU=19.818,t60IU=24.833,P＜0.05;MDA t10IU=5.345,t20IU=10.278,t40It/=18.571,t60IU=20.247,P＜0.05);negative correlation with the relation rate of LDH and the content of MDA(r=-0.976,P=0.024;r=the content of MDA(r=-0.954,P=0.046). Conclusion EPO can protect hFRPE ceils from the injuryof H2O2,the mechanism may be related to the activation of NF-KB.