促红细胞生成素对人视网膜色素上皮细胞氧化损伤的保护作用

Protective effects of erythropoietin on human retinal pigment epithelial cells injured by hydrogen peroxide

目的探讨促红细胞生成素对过氧化氢氧化损伤人视网膜色素上皮细胞的保护作用及其机制。方法采用600μmol.L-1的过氧化氢造成体外培养的人视网膜色素上皮细胞氧化损伤模型。实验设计分为5组:对照组,过氧化氢模型组,过氧化氢+10 kIU.L-1EPO组,过氧化氢+20 kIU.L-1EPO组和过氧化氢+40 kIU.L-1EPO组。MTT法检测细胞活性,测定细胞内超氧化物歧化酶(SOD)和丙二醛(MDA)含量,Annexin V-FITC/PI双染色流式细胞法测定细胞的凋亡率。结果过氧化氢模型组RPE细胞的活性明显降低,不同浓度的EPO药物干预组的细胞活性较模型组明显升高,并随着EPO浓度的升高而升高。过氧化氢模型组RPE细胞SOD活性降低,MDA的含量增加;而EPO药物干预组SOD活性较模型组明显升高,MDA含量减少,差异有显著性。过氧化氢模型组RPE细胞的凋亡率升高,而不同浓度的EPO药物干预组降低RPE的凋亡率,凋亡率随着EPO浓度的升高而降低。结论促红细胞生成素对过氧化氢诱导的人RPE细胞的氧化损伤有保护作用,其机制可能与抑制氧化反应、提高抗氧化酶活性、减少细胞凋亡有关。

Aim To explore the protective effects of erythropoietin （EPO） on human retinal pigment epithelial （RPE） cells injured by hydrogen peroxide （H2O2 ）. Methods RPE cells were randomly divided into five groups： control, H2O2 model, H202 ＋ 10 kIU· L^-1 EPO, H202 ＋20 kIU ·L^-1 EPO, H2O2 ＋40 kIU · L^-1 EPO group. MTT method was used to detect the viability of human RPE cells. The content of malondialdehyde （MDA） and the activity of superoxide dimutase （SOD） of the RPE cells were examined. The effect of EPO on H2O2-induced RPE cells apoptosis was assessed by Annexin V-fluorescein isothiocyanate / Propidium iodium（ Annexin V-FITC/PI ） staining. Results EPO greatly enhanced the viability of human RPE cells injured by H2O2. EPO markedly promoted the viability of SOD and decrease MDA in RPE cells injured by H2O2. Apoptosis of RPE cells occurs in H2O2 model group. But in EPO treatment groups the apoptosis rate of RPE cells was decreased. Conclusion EPO rescues human RPE cells from oxidative injury induced by H2O2, which may inhibit apoptosis by promoting the viability of SOD and decreasing MDA. EPO has protective effect on human RPE oxidative injury.